TY - JOUR AB - Antinuclear antibodies (ANAs) are central biomarkers in rheumatological conditions and can drive disease pathology. Much less is known about the role of ANAs in neurological symptoms, although a number of experimental studies have demonstrated direct effects on neuronal function, for example in neuropsychiatric lupus erythematosus. Moreover, it is unclear whether the ANAs detected in HEp-2 cell-based assays, the gold standard for ANA diagnostics, can also be recognized in modern screening assays for anti-neuronal autoimmunity, such as staining on rodent brain sections or neuronal cultures. In this study, we therefore conducted a comparative mapping of ANA-positive sera with well-characterized HEp-2 patterns to central nervous system (CNS) tissue, utilizing fixed and unfixed murine brain sections and primary murine neurons. We screened 74 ANA-positive sera classified into 14 individual patterns and combinations thereof. Majority of the samples reacted with fixed primary neurons (99%, 73/74 sera), followed by fixed brain sections (93%, 69/74), but much less to unfixed mouse brain (54%, 40/74). While the PM/SCL- and RPOI-positive sera showed no binding to unfixed brain sections, the U1RNP (U1 nuclear ribonucleoprotein particle) and FBLN (fibrillarin) ANAs reacted strongly across all assays, indicating differences in antigen accessibility. These findings suggest that the majority of ANAs can interact with neural components, which may obscure the detection of other anti-neuronal autoantibodies. The foundational mapping of ANA binding in CNS tissue provided here can also facilitate recognition of "CNS-specific ANAs," which bind to neuronal autoantigens but not to HEp-2 cells. Future studies should explore the association with certain neurological manifestations and the role of ANAs in neuronal pathology. AU - Li, L.Y.* AU - Höltje, M.* AU - Rasmussen, H.F.* AU - Halle, L. AU - Mayrhofer, M. AU - Blüthner, M.* AU - Prüss, H.* C1 - 75866 C2 - 58162 TI - Binding of established antinuclear antibodies to neurons depends on tissue fixation and underlying autoantigens. JO - Front. Immunol. VL - 16 PY - 2025 SN - 1664-3224 ER - TY - JOUR AB - BACKGROUND: Sex, race and geographic location may affect vaccine-induced immune responses, yet few preventive HIV vaccine trials have systematically evaluated such effects. The main objective of this study was therefore to examine the role of these factors on vaccine-induced HIV-specific immune responses within the HVTN 204 trial. This randomized, double-blinded, placebo-controlled phase 2 trial enrolled 480 Black and Caucasian adults from Africa and the Americas, who received a trivalent DNA-HIV-1 vaccine prime followed by a rAd5 vector HIV-1 vaccine boost. METHODS: Available serum samples from baseline and four weeks after the final vaccination boost from Black (n=85, 59% female) and Caucasian (n=49, 51% female) HVTN 204 vaccine recipients from South Africa and the United States of America were studied using an Enzyme-Linked Immunosorbent Assay to determine titers of Envelope-specific IgG1, IgG3 and IgA antibodies. Recognition of linear Envelope peptide-specific IgG responses was mapped in a randomly selected subgroup analysis using a custom-designed peptide microarray (n = 41, 49% female). Associations between vaccine-induced Envelope-specific antibody responses and sex assigned at birth (female or male), race and geographic location were then analyzed by the Mann-Whitney U test, Fisher's exact test and multivariate logistic regression. RESULTS: Four weeks post-final vaccination boost, we observed that Envelope-specific antibody titers were significantly increased for IgG1 but reduced for IgA in females (female vs. male median titer: 900 vs. 300, p=0.030 and <100 vs. 100, p=0.007, respectively). Multivariate logistic regression confirmed that female sex increased the odds for higher Envelope-specific IgG1 and low IgA titers compared to males. In terms of antibody epitopes, the V2 region was more frequently recognized in females than males (p=0.008). Race and geographic location had no apparent influence on antibody isotype titers investigated. CONCLUSION: Female sex was associated with higher vaccine-induced IgG Envelope-specific binding antibody titers and recognition of V2 region of HIV Envelope in HVTN 204 volunteers. No such associations were detected for race or geographic location. Understanding biological factors driving these sex-based differences may improve the design of a new generation of HIV vaccine candidates. AU - Mbuya, W.* AU - Horvath, A.* AU - Held, K. AU - Maganga, L.* AU - Hoelscher, M. AU - Bekker, L.G.* AU - Duerr, A.* AU - Moodie, Z.* AU - Churchyard, G.* AU - Keefer, M.C.* AU - Viegas, E.* AU - Moog, C.* AU - Geldmacher, C. AU - Chachage, M.* C1 - 75742 C2 - 58155 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Effects of sex but not race and geographic origin on vaccine-induced HIV-specific antibody responses. JO - Front. Immunol. VL - 16 PB - Frontiers Media Sa PY - 2025 SN - 1664-3224 ER - TY - JOUR AB - The human genome contains ~8% of endogenous retroviruses (HERVs), whose reactivation has been implicated in diseases such as cancer and autoimmune disorders. Among these, HERV-K10 has attracted attention for its potential role in immune modulation and viral infections. This study investigates HERV-K10 expression in hepatitis virus infections, focusing on its impact on host gene expression and immune responses. We analyzed HERV-K10 in PBMCs from patients chronically infected with hepatitis C virus (HCV) and in HBV-infected liver cell models. Our results show a significant upregulation of HERV-K10 in HBV-infected HepG2-NTCP cells, HCV-infected PBMCs, and a trend in HBV-infected primary hepatocytes. HERV-K10 activation was specific to hepatitis infection, as no effect was seen with HBV entry inhibitors, adenovirus 5 infection or infection with other RNA viruses. RNA sequencing of HBV-infected HepG2-NTCP cells revealed distinct clustering based on HERV expression profiles, including HERV-K10 encoding the MAG1 domain, an immune response target. To investigate the potential immunomodulatory role of HERV-K10 MAG1, we vaccinated mice with the MAG1 peptide, which resulted in activation of CD4+ and CD8+ T-cell responses and higher levels of MAG1-specific antibodies. Furthermore, chronic hepatitis B patients exhibited an immune response to MAG1 characterized by elevated levels of Interleukin-6 (IL-6) and interleukin-1β (IL-1β) cytokines. Taken together, our data suggest that HERV-K10 plays an important role in immune modulation during viral hepatitis infection and may contribute to the pathogenesis of autoimmune diseases. AU - Özer, S. AU - Strobelt, R. AU - Kosinska, A. AU - Frishman, G. AU - Wettengel, J.M. AU - Pleninger, L. AU - Körber, N. AU - Liang, W.* AU - Ates Öz, E. AU - Zuniga, M. AU - Bauer, T. AU - Ebert, G. AU - Protzer, U. AU - Vincendeau, M. C1 - 75698 C2 - 57926 TI - HERV-K10 as a mediator of immune modulation in hepatitis infections. JO - Front. Immunol. VL - 16 PY - 2025 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Women with a history of gestational diabetes mellitus (GDM) are at high risk of developing prediabetes or type 2 diabetes later in life. Recent studies have highlighted the regulation and function of innate lymphoid cells (ILCs) in metabolic homeostasis. However, the multifactorial impact of both overweight/obesity and GDM on the immunological profile of circulating ILCs and the progression to prediabetes are not yet fully elucidated. METHODS: Blood samples from 42 women with a history of insulin-treated GDM (GDMi), 33 women with a history of GDM without insulin treatment during pregnancy (GDM), and 45 women after a normoglycemic pregnancy (Ctrl) participating in the ongoing observational PPSDiab study were analyzed by flow cytometry for markers of ILC subsets at the baseline visit (3-16 months postpartum; Visit 1) and 5 years postpartum (58-66 months postpartum; Visit 2). RESULTS: During the first 5 years postpartum, 18 women of the GDMi group (42.8%), 10 women of the GDM group (30.3%), and 8 participants of the Ctrl group (17.8%) developed prediabetes, respectively. Total circulating type 1 innate lymphoid cells (ILC1s) and NK cell numbers as well as percent HLA-DR+ ILC1s were increased in GDMi versus GDM and Ctrl women both at the baseline visit and the 5-year follow-up. Although ILC subsets at Visit 1 could not predict the progression from GDM to prediabetes, ILC2 frequency was associated with insulin sensitivity index (ISI), whereas percent HLA-DR+ ILC1s were inversely correlated. Moreover, circulating leukocytes and total NK cells were associated with waist circumference and fat mass both at Visit 1 and Visit 2. DISCUSSION: Our findings introduce human ILCs as a potential therapeutic target deserving further exploration. TRIAL REGISTRATION: Study ID 300-11. AU - Sbierski-Kind, J. AU - Schlickeiser, S.* AU - Semeia, L. AU - Harada, S.* AU - Pappa, E.* AU - Cujar, J.V.* AU - Katschke, M.T.* AU - Gar, C.* AU - Lechner, A.* AU - Birkenfeld, A.L. AU - Ferrari, U.* AU - Seissler, J.* C1 - 73745 C2 - 57210 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Association of overweight/obesity and insulin resistance with activation of circulating innate lymphoid cells in women after gestational diabetes mellitus. JO - Front. Immunol. VL - 16 PB - Frontiers Media Sa PY - 2025 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Treating autoimmune diseases without nonspecific immunosuppression remains challenging. To prevent or treat these conditions through targeted immunotherapy, we developed a clinical-stage nanoparticle platform that leverages the tolerogenic capacity of liver sinusoidal endothelial cells (LSECs) to restore antigen-specific immune tolerance. METHODS: In vivo efficacy was evaluated in various CD4+ T cell-mediated disease models, including preventive and therapeutic models of myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis (EAE), ovalbumin-sensitized delayed-type hypersensitivity (DTH), and the spontaneous type 1 diabetes model. Nanoparticle-induced antigen-specific immune responses were also analyzed through adoptive transfers of 2D2 transgenic T cells into wild-type mice, followed by nanoparticle administration. RESULTS: The peptide-conjugated nanoparticles displayed a uniform size distribution (25-30 nm). Their coupling efficiency for peptides with unfavorable physicochemical properties was significantly enhanced by a proprietary linker technology. Preferential LSEC targeting of nanoparticles coupled with fluorescently labeled peptides was confirmed via intravital microscopy and flow cytometry. Intravenous nanoparticle administration significantly reduced disease severity and demyelination in EAE, independent of prednisone at maintenance doses, and suppressed target tissue inflammation in the DTH model. Furthermore, prophylactic administration of a mixture of nanoparticles coupled with five autoantigenic peptides significantly lowered the hyperglycemia incidence of the non-obese diabetic mice. Mechanistically, the tolerizing effects were associated with the induction of antigen-specific regulatory T cells and T cell anergy, which counteract proinflammatory T cells in the target tissue. CONCLUSION: Our findings demonstrate that peptide-loaded nanoparticles preferentially deliver disease-relevant peptides to LSECs, thereby inducing antigen-specific immune tolerance. This versatile clinical-stage nanoparticle platform holds promise for clinical application across multiple autoimmune diseases. AU - Wang, S.H.* AU - Serr, I. AU - Digigow, R.* AU - Metzler, B.* AU - Surnov, A. AU - Gottwick, C.* AU - Alsamman, M.* AU - Krzikalla, D.* AU - Heine, M.* AU - Zahlten, M.* AU - Widera, A.* AU - Mungalpara, D.* AU - Şeleci, M.* AU - Fanzutti, M.* AU - Marques Mesquita, L.M.* AU - Vocaturo, A.L.* AU - Herkel, J.* AU - Carambia, A.* AU - Schröter, C.* AU - Sarko, D.* AU - Pohlner, J.* AU - Daniel, C. AU - de Min, C.* AU - Fleischer, S.* C1 - 73919 C2 - 57241 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Nanoparticle platform preferentially targeting liver sinusoidal endothelial cells induces tolerance in CD4+ T cell-mediated disease models. JO - Front. Immunol. VL - 16 PB - Frontiers Media Sa PY - 2025 SN - 1664-3224 ER - TY - JOUR AB - BACKGROUND: Allergen-specific immunotherapy (AIT) is able to restore immune tolerance to allergens in allergic patients. However, some patients do not or only poorly respond to current treatment protocols. Therefore, there is a need for deeper mechanistic insights and further improvement of treatment strategies. The relevance of the aryl hydrocarbon receptor (AhR), a ligand-dependent transcription factor, has been investigated in several inflammatory diseases, including allergic asthma. However, its potential role in AIT still needs to be addressed. METHODS: A murine model of AIT in ovalbumin-induced allergic airway inflammation was performed in AhR-deficient (AhR-/-) and wild-type mice. Furthermore, AIT was combined with the application of the high-affinity AhR agonist 10-chloro-7H-benzimidazo[2,1-a]benzo[de]iso-quinolin-7-one (10-Cl-BBQ) as an adjuvant to investigate the effects of AhR activation on therapeutic outcome. RESULTS: Although AhR-/- mice suffer stronger allergic responses than wild-type mice, experimental AIT is comparably effective in both. Nevertheless, combining AIT with the administration of 10-Cl-BBQ improved therapeutic effects by an AhR-dependent mechanism, resulting in decreased cell counts in the bronchoalveolar fluid, decreased pulmonary Th2 and Th17 cell levels, and lower sIgE levels. CONCLUSION: This study demonstrates that the success of AIT is not dependent on the AhR. However, targeting the AhR during AIT can help to dampen inflammation and improve tolerogenic vaccination. Therefore, AhR ligands might represent promising candidates as immunomodulators to enhance the efficacy of AIT. AU - Heine, S. AU - Alessandrini, F. AU - Grosch, J. AU - Graß, C. AU - Heldner, A. AU - Schnautz, B. AU - Buters, J.T.M. AU - Slusarenko, B.O. AU - Krappmann, D. AU - Fallarino, F.* AU - Ohnmacht, C. AU - Schmidt-Weber, C.B. AU - Blank, S. C1 - 70901 C2 - 55974 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland SP - 1397072 TI - Activation of the aryl hydrocarbon receptor improves allergen-specific immunotherapy of murine allergic airway inflammation: A novel adjuvant option? JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - CARD-BCL10-MALT1 (CBM) signalosomes connect distal signaling of innate and adaptive immune receptors to proximal signaling pathways and immune activation. Four CARD scaffold proteins (CARD9, 10, 11, 14) can form seeds that nucleate the assembly of BCL10-MALT1 filaments in a cell- and stimulus-specific manner. MALT1 (also known as PCASP1) serves a dual function within the assembled CBM complexes. By recruiting TRAF6, MALT1 acts as a molecular scaffold that initiates IκB kinase (IKK)/NF-κB and c-Jun N-terminal kinase (JNK)/AP-1 signaling. In parallel, proximity-induced dimerization of the paracaspase domain activates the MALT1 protease which exerts its function by cleaving a set of specific substrates. While complete MALT1 ablation leads to immune deficiency, selective destruction of either scaffolding or protease function provokes autoimmune inflammation. Thus, balanced MALT1-TRAF6 recruitment and MALT1 substrate cleavage are critical to maintain immune homeostasis and to promote optimal immune activation. Further, MALT1 protease activity drives the survival of aggressive lymphomas and other non-hematologic solid cancers. However, little is known about the relevance of the cleavage of individual substrates for the pathophysiological functions of MALT1. Unbiased serendipity, screening and computational predictions have identified and validated ~20 substrates, indicating that MALT1 targets a quite distinct set of proteins. Known substrates are involved in CBM auto-regulation (MALT1, BCL10 and CARD10), regulation of signaling and adhesion (A20, CYLD, HOIL-1 and Tensin-3), or transcription (RelB) and mRNA stability/translation (Regnase-1, Roquin-1/2 and N4BP1), indicating that MALT1 often targets multiple proteins involved in similar cellular processes. Here, we will summarize what is known about the fate and functions of individual MALT1 substrates and how their cleavage contributes to the biological functions of the MALT1 protease. We will outline what is needed to better connect critical pathophysiological roles of the MALT1 protease with the cleavage of distinct substrates. AU - Nemati Moud, B. AU - Ober, F. AU - O'Neill, T.J. AU - Krappmann, D. C1 - 70828 C2 - 55757 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - MALT1 substrate cleavage: What is it good for? JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - BACKGROUND: High dietary sodium intake is a major cardiovascular risk factor and adversely affects blood pressure control. Patients with primary aldosteronism (PA) are at increased cardiovascular risk, even after medical treatment, and high dietary sodium intake is common in these patients. Here, we analyze the impact of a moderate dietary sodium restriction on microbiome composition and immunophenotype in patients with PA. METHODS: Prospective two-stage clinical trial including two subgroups: 15 treatment-naive PA patients compared to matched normotensive controls; and 31 PA patients on mineralocorticoid receptor antagonist treatment before and three months after sodium restriction. Patients underwent blood pressure measurements, laboratory tests, analysis of peripheral blood mononuclear cells via flow cytometry and microbiome analysis. RESULTS: We observed a higher percentage of Tregs in treatment-naive PA patients (p = 0.0303), while the abundance of Bacteroides uniformis was higher in PA patients compared to normotensive controls (p = 0.00027) and the abundance of Lactobacillus species however was higher in the subgroup of normotensive controls (p = 0.0290). Sodium restriction was accompanied by a decrease in pro-inflammatory Tc17 cells in male patients (p = 0.0081, females p = 0.3274). Bacteroides uniformis abundance was higher in female patients (0.01230, p = 0.0016) and decreased upon sodium restriction (0.002309, p = 0.0068). CONCLUSION: Dietary sodium restriction in patients with PA modulates the peripheral immune cell composition toward a less inflammatory phenotype. This suggests a potential mechanism by which sodium reduction modulates immune cell composition, leading to blood pressure reduction and positively impacting cardiovascular risk. AU - Nowotny, H.F.* AU - Zheng, T.* AU - Seiter, T.M.* AU - Ju, J.* AU - Schneider, H.* AU - Kroiss, M.* AU - Sarkis, A.L.* AU - Sturm, L.* AU - Britz, V.* AU - Lechner, A. AU - Potzel, A. AU - Kunz, S.* AU - Bidlingmaier, M.* AU - Neuhaus, K.* AU - Gottschlich, A.* AU - Kobold, S. AU - Reisch, N.* AU - Schirmer, M.* AU - Reincke, M.* AU - Adolf, C.* C1 - 72929 C2 - 56791 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Sex-dependent modulation of T and NK cells and gut microbiome by low sodium diet in patients with primary aldosteronism. JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Obesity is associated with a plethora of health complications, including increased susceptibility to infections or decreased vaccine efficacy, partly due to dysregulated immune responses. Monocytes play a crucial role in innate immunity, yet their functional alterations in obesity remain poorly understood. METHODS: Here, we employed proteomic and metabolomic analyses to investigate monocyte characteristics in individuals with overweight, obesity, impaired glucose tolerance (IGT), and type 2 diabetes (T2D), compared to lean donors. RESULTS AND DISCUSSION: Our results revealed distinct molecular signatures in monocytes from individuals with obesity, with significant alterations in pathways related to metabolism, cellular migration, and phagocytosis. Moreover, LPS-induced activation of monocytes unveiled heightened metabolic reprogramming towards glycolysis in subjects with obesity accompanied by dysregulated cytokine responses and elevated oxidative stress. Additionally, monocytes from donors with obesity exhibited increased lipid droplet accumulation. These findings shed light on the immunometabolic dysregulation underlying obesity-associated immune dysfunction, highlighting potential targets for therapeutic intervention. AU - Radushev, V.* AU - Karkossa, I.* AU - Berg, J.* AU - von Bergen, M.* AU - Engelmann, B.* AU - Rolle-Kampczyk, U.* AU - Blüher, M. AU - Wagner, U.* AU - Schubert, K.* AU - Rossol, M.* C1 - 71299 C2 - 56032 TI - Dysregulated cytokine and oxidative response in hyper-glycolytic monocytes in obesity. JO - Front. Immunol. VL - 15 PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - To design new CARs targeting hepatitis B virus (HBV), we isolated human monoclonal antibodies recognizing the HBV envelope proteins from single B cells of a patient with a resolved infection. HBV-specific memory B cells were isolated by incubating peripheral blood mononuclear cells with biotinylated hepatitis B surface antigen (HBsAg), followed by single-cell flow cytometry-based sorting of live, CD19+ IgG+ HBsAg+ cells. Amplification and sequencing of immunoglobulin genes from single memory B cells identified variable heavy and light chain sequences. Corresponding immunoglobulin chains were cloned into IgG1 expression vectors and expressed in mammalian cells. Two antibodies named 4D06 and 4D08 were found to be highly specific for HBsAg, recognized a conformational and a linear epitope, respectively, and showed broad reactivity and neutralization capacity against all major HBV genotypes. 4D06 and 4D08 variable chain fragments were cloned into a 2nd generation CAR format with CD28 and CD3zeta intracellular signaling domains. The new CAR constructs displayed a high functional avidity when expressed on primary human T cells. CAR-grafted T cells proved to be polyfunctional regarding cytokine secretion and killed HBV-positive target cells. Interestingly, background activation of the 4D08-CAR recognizing a linear instead of a conformational epitope was consistently low. In a preclinical model of chronic HBV infection, murine T cells grafted with the 4D06 and the 4D08 CAR showed on target activity indicated by a transient increase in serum transaminases, and a lower number of HBV-positive hepatocytes in the mice treated. This study demonstrates an efficient and fast approach to identifying pathogen-specific monoclonal human antibodies from small donor cell numbers for the subsequent generation of new CARs. AU - Schreiber, S. AU - Dressler, L.S. AU - Loffredo-Verde, E. AU - Asen, T. AU - Färber, S. AU - Wang, W.* AU - Groll, T.* AU - Chakraborty, A. AU - Kolbe, F. AU - Kreer, C.* AU - Kosinska, A. AU - Simon, S.* AU - Urban, S.* AU - Klein, F.* AU - Riddell, S.R.* AU - Protzer, U. C1 - 70619 C2 - 55773 TI - CARs derived from broadly neutralizing, human monoclonal antibodies identified by single B cell sorting target hepatitis B virus-positive cells. JO - Front. Immunol. VL - 15 PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - Sebaceous glands drive acne, however, their role in other inflammatory skin diseases remains unclear. To shed light on their potential contribution to disease development, we investigated the spatial transcriptome of sebaceous glands in psoriasis and atopic dermatitis patients across lesional and non-lesional human skin samples. Both atopic dermatitis and psoriasis sebaceous glands expressed genes encoding key proteins for lipid metabolism and transport such as ALOX15B, APOC1, FABP7, FADS1/2, FASN, PPARG, and RARRES1. Also, inflammation-related SAA1 was identified as a common spatially variable gene. In atopic dermatitis, genes mainly related to lipid metabolism (e.g. ACAD8, FADS6, or EBP) as well as disease-specific genes, i.e., Th2 inflammation-related lipid-regulating HSD3B1 were differentially expressed. On the contrary, in psoriasis, more inflammation-related spatially variable genes (e.g. SERPINF1, FKBP5, IFIT1/3, DDX58) were identified. Other psoriasis-specific enriched pathways included lipid metabolism (e.g. ACOT4, S1PR3), keratinization (e.g. LCE5A, KRT5/7/16), neutrophil degranulation, and antimicrobial peptides (e.g. LTF, DEFB4A, S100A7-9). In conclusion, our results show that sebaceous glands contribute to skin homeostasis with a cell type-specific lipid metabolism, which is influenced by the inflammatory microenvironment. These findings further support that sebaceous glands are not bystanders in inflammatory skin diseases, but can actively and differentially modulate inflammation in a disease-specific manner. AU - Seiringer, P.* AU - Hillig, C. AU - Schäbitz, A.* AU - Jargosch, M. AU - Pilz, A.C.* AU - Eyerich, S. AU - Szegedi, A.* AU - Sochorová, M.* AU - Gruber, F.* AU - Zouboulis, C.C.* AU - Biedermann, T.* AU - Menden, M.P. AU - Eyerich, K.* AU - Törőcsik, D.* C1 - 70088 C2 - 55413 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Spatial transcriptomics reveals altered lipid metabolism and inflammation-related gene expression of sebaceous glands in psoriasis and atopic dermatitis. JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: P2X receptors are a family of homo- and heterotrimeric cation channels gated by extracellular ATP. The P2X4 and P2X7 subunits show overlapping expression patterns and have been involved in similar physiological processes, such as pain and inflammation as well as various immune cell functions. While formation of P2X2/P2X3 heterotrimers produces a distinct pharmacological phenotype and has been well established, functional identification of a P2X4/P2X7 heteromer has been difficult and evidence for and against a physical association has been found. Most of this evidence stems, however, from in vitro model systems. METHODS: Here, we used a P2X7-EGFP BAC transgenic mouse model as well as P2X4 and P2X7 knock-out mice to re-investigate a P2X4-P2X7 interaction in mouse lung by biochemical and immunohistochemical experiments as well as quantitative expression analysis. RESULTS: No detectable amounts of P2X4 could be co-purified from mouse lung via P2X7-EGFP. In agreement with these findings, immuno-histochemical analysis using a P2X7-specific nanobody revealed only limited overlap in the cellular and subcellular localizations of P2X4 and P2X7 in both the native lung tissue and primary cells. Comparison of P2X4 and P2X7 transcript and protein levels in the respective gene-deficient and wild type mice showed no mutual interrelation between their expression levels in whole lungs. However, a significantly reduced P2rx7 expression was found in alveolar macrophages of P2rx4 -/- mice. DISCUSSION: In summary, our detailed analysis of the cellular and subcellular P2X4 and P2X7 localization and expression does not support a physiologically relevant direct association of P2X4 and P2X7 subunits or receptors in vivo. AU - Sierra-Marquez, J.* AU - Schaller, L.* AU - Sassenbach, L.* AU - Ramírez-Fernández, A.* AU - Alt, P.* AU - Rissiek, B.* AU - Zimmer, B.* AU - Schredelseker, J.* AU - Hector, J.* AU - Stähler, T.* AU - Koch-Nolte, F.* AU - Staab-Weijnitz, C.A. AU - Dietrich, A.* AU - Kopp, R.* AU - Nicke, A.* C1 - 71027 C2 - 55989 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Different localization of P2X4 and P2X7 receptors in native mouse lung - lack of evidence for a direct P2X4-P2X7 receptor interaction. JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - The mucosal immunity is crucial for restricting SARS-CoV-2 at its entry site. Intramuscularly applied vaccines against SARS-CoV-2 stimulate high levels of neutralizing Abs in serum, but the impact of these intramuscular vaccinations on features of mucosal immunity is less clear. Here, we analyzed kinetic and functional properties of anti-SARS-CoV-2 Abs in the saliva after vaccination with BNT162b2. We analyzed a total of 24 healthy donors longitudinally for up to 16 months. We found that specific IgG appeared in the saliva after the second vaccination, declined thereafter and reappeared after the third vaccination. Adjusting serum and saliva for the same IgG concentration revealed a strong correlation between the reactivity in these two compartments. Reactivity to VoCs correlated strongly as seen by ELISAs against RBD variants and by live-virus neutralizing assays against replication-competent viruses. For further functional analysis, we purified IgG and IgA from serum and saliva. In vaccinated donors we found neutralizing activity towards authentic virus in the IgG, but not in the IgA fraction of the saliva. In contrast, IgA with neutralizing activity appeared in the saliva only after breakthrough infection. In serum, we found neutralizing activity in both the IgA and IgG fractions. Together, we show that intramuscular mRNA vaccination transiently induces a mucosal immunity that is mediated by IgG and thus differs from the mucosal immunity after infection. Waning of specific mucosal IgG might be linked to susceptibility for breakthrough infection. AU - Winklmeier, S.* AU - Rübsamen, H.* AU - Ozdemir, C.* AU - Wratil, P.R.* AU - Lupoli, G.* AU - Stern, M.* AU - Schneider, C.* AU - Eisenhut, K.* AU - Ho, S.* AU - Wong, H.K.* AU - Taskin, D.* AU - Petry, M.* AU - Weigand, M.* AU - Eichhorn, P.* AU - Fösel, B. AU - Mader, S.* AU - Keppler, O.T.* AU - Kümpfel, T.* AU - Meinl, E.* C1 - 70059 C2 - 55389 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Intramuscular vaccination against SARS-CoV-2 transiently induces neutralizing IgG rather than IgA in the saliva. JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AU - Zhang, B.* AU - Mautner, J. AU - Münz, C.* C1 - 71279 C2 - 55998 TI - Editorial: EBV-induced T cell immunity in cancers. JO - Front. Immunol. VL - 15 PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - Myelin sheath, as the multilayer dense structure enclosing axons in humans and other higher organisms, may rupture due to various injury factors after spinal cord injury, thus producing myelin debris. The myelin debris contains a variety of myelin-associated inhibitors (MAIs) and lipid, all inhibiting the repair after spinal cord injury. Through summary and analysis, the present authors found that the inhibition of myelin debris can be mainly divided into two categories: firstly, the direct inhibition mediated by MAIs; secondly, the indirect inhibition mediated by lipid such as cholesterol. It is worth noting that phagocytes are required in the latter indirect inhibition, such as professional phagocytes (macrophages et al.) and non-professional phagocytes (astrocytes et al.). Moreover, complement and the immune system also participate in the phagocytosis of myelin debris, working together with phagocytes to aggravate spinal cord injury. In conclusion, this paper focuses on the direct and indirect effects of myelin debris on spinal cord injury, aiming to provide new inspiration and reflection for the basic research of spinal cord injury and the conception of related treatment. AU - Zhou, Y.* AU - Xu, T.* AU - Han, W. AU - Wu, Z.* AU - Yang, C.* AU - Chen, X.* C1 - 72693 C2 - 56695 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - A review focuses on a neglected and controversial component of SCI: Myelin debris. JO - Front. Immunol. VL - 15 PB - Frontiers Media Sa PY - 2024 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Mesenchymal stem cells (MSCs) are considered to be the most promising stem cell type for cell-based therapies in regenerative medicine. Based on their potential to home to diseased body sites following a therapeutically application, these cells could (i) differentiate then into organ-specific cell types to locally restore injured cells or, most prominently, (ii) foster tissue regeneration including immune modulations more indirectly by secretion of protective growth factors and cytokines. As tissue-resident stem cells of mesenchymal origin, these cells are morphologically and even molecularly- at least concerning the classical marker genes- indistinguishable from similar lineage cells, particularly fibroblasts. METHODS: Here we used microarray-based gene expression and global DNA methylation analyses as well as accompanying computational tools in order to specify differences between MSCs and fibroblasts, to further unravel potential identity genes and to highlight MSC signaling pathways with regard to their trophic and immunosuppressive action. RESULTS: We identified 1352 differentially expressed genes, of which in the MSCs there is a strong signature for e.g., KRAS signaling, known to play essential role in stemness maintenance, regulation of coagulation and complement being decisive for resolving inflammatory processes, as well as of wound healing particularly important for their regenerative capacity. Genes upregulated in fibroblasts addressed predominately transcription and biosynthetic processes and mapped morphological features of the tissue. Concerning the cellular identity, we specified the already known HOX code for MSCs, established a potential HOX code for fibroblasts, and linked certain HOX genes to functional cell-type-specific properties. Accompanied methylation profiles revealed numerous regions, especially in HOX genes, being differentially methylated, which might provide additional biomarker potential. DISCUSSION: Conclusively, transcriptomic together with epigenetic signatures can be successfully be used for the definition (cellular identity) of MSCs versus fibroblasts as well as for the determination of the superior functional properties of MSCs, such as their immunomodulatory potential. AU - Budeus, B.* AU - Unger, K. AU - Hess J. AU - Sentek, H.* AU - Klein, D.* C1 - 68665 C2 - 54870 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Comparative computational analysis to distinguish mesenchymal stem cells from fibroblasts. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Allogeneic stem cell transplantation is used to cure hematologic malignancies or deficiencies of the hematopoietic system. It is associated with severe immunodeficiency of the host early after transplant and therefore early reactivation of latent herpesviruses such as CMV and EBV within the first 100 days are frequent. Small studies and case series indicated that application of herpes virus specific T cells can control and prevent disease in this patient population. METHODS: We report the results of a randomized controlled multi centre phase I/IIa study (MULTIVIR-01) using a newly developed T cell product with specificity for CMV and EBV derived from the allogeneic stem cell grafts used for transplantation. The study aimed at prevention and preemptive treatment of both viruses in patients after allogeneic stem cell transplantation targeting first infusion on day +30. Primary endpoints were acute transfusion reaction and acute-graft versus-host-disease after infusion of activated T cells. RESULTS: Thirty-three patients were screened and 9 patients were treated with a total of 25 doses of the T cell product. We show that central manufacturing can be achieved successfully under study conditions and the product can be applied without major side effects. Overall survival, transplant related mortality, cumulative incidence of graft versus host disease and number of severe adverse events were not different between treatment and control groups. Expansion of CMV/EBV specific T cells was observed in a fraction of patients, but overall there was no difference in virus reactivation. DISCUSSION: Our study results indicate peptide stimulated epitope specific T cells derived from stem cell grafts can be administered safely for prevention and preemptive treatment of reactivation without evidence for induction of acute graft versus host disease. CLINICAL TRIAL REGISTRATION: https://clinicaltrials.gov, identifier NCT02227641. AU - Gerbitz, A.* AU - Gary, R.* AU - Aigner, M.* AU - Moosmann, A. AU - Kremer, A.* AU - Schmid, C.* AU - Hirschbuehl, K.* AU - Wagner, E.* AU - Hauptrock, B.* AU - Teschner, D.* AU - Roesler, W.* AU - Spriewald, B.* AU - Tischer, J.* AU - Moi, S.* AU - Balzer, H.* AU - Schaffer, S.* AU - Bausenwein, J.* AU - Wagner, A.* AU - Schmidt, F.* AU - Brestrich, J.* AU - Ullrich, B.* AU - Maas, S.* AU - Herold, S.* AU - Strobel, J.* AU - Zimmermann, R.* AU - Weisbach, V.* AU - Hansmann, L.* AU - Lammoglia-Cobo, F.* AU - Remberger, M.* AU - Stelljes, M.* AU - Ayuk, F.* AU - Zeiser, R.* AU - Mackensen, A.* C1 - 68775 C2 - 54985 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Prevention of CMV/EBV reactivation by double-specific T cells in patients after allogeneic stem cell transplantation: Results from the randomized phase I/IIa MULTIVIR-01 study. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AU - Kapellos, T. AU - Nawijn, M.C.* C1 - 67860 C2 - 54338 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Editorial: Molecular mechanisms regulating phenotypic heterogeneity in human inflammatory diseases. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Interstitial lung disease (ILD) is a heterogenous group of lung disorders where destruction and incomplete regeneration of the lung parenchyma often results in persistent architectural distortion of the pulmonary scaffold. Continuous mesenchyme-centered, disease-relevant signaling likely initiates and perpetuates the fibrotic remodeling process, specifically targeting the epithelial cell compartment, thereby destroying the gas exchange area. METHODS: With the aim of identifying functional mediators of the lung mesenchymal-epithelial crosstalk with potential as new targets for therapeutic strategies, we developed a 3D organoid co-culture model based on human induced pluripotent stem cell-derived alveolar epithelial type 2 cells that form alveolar organoids in presence of lung fibroblasts from fibrotic-ILD patients, in our study referring to cases of pulmonary fibrosis, as well as control cell line (IMR-90). RESULTS: While organoid formation capacity and size was comparable in the presence of fibrotic-ILD or control lung fibroblasts, metabolic activity was significantly increased in fibrotic-ILD co-cultures. Alveolar organoids cultured with fibrotic-ILD fibroblasts further demonstrated reduced stem cell function as reflected by reduced Surfactant Protein C gene expression together with an aberrant basaloid-prone differentiation program indicated by elevated Cadherin 2, Bone Morphogenic Protein 4 and Vimentin transcription. To screen for key mediators of the misguided mesenchymal-to-epithelial crosstalk with a focus on disease-relevant inflammatory processes, we used mass spectrometry and characterized the secretome of end stage fibrotic-ILD lung fibroblasts in comparison to non-chronic lung disease (CLD) patient fibroblasts. Out of the over 2000 proteins detected by this experimental approach, 47 proteins were differentially abundant comparing fibrotic-ILD and non-CLD fibroblast secretome. The fibrotic-ILD secretome profile was dominated by chemokines, including CXCL1, CXCL3, and CXCL8, interfering with growth factor signaling orchestrated by Interleukin 11 (IL11), steering fibrogenic cell-cell communication, and proteins regulating extracellular matrix remodeling including epithelial-to-mesenchymal transition. When in turn treating alveolar organoids with IL11, we recapitulated the co-culture results obtained with primary fibrotic-ILD fibroblasts including changes in metabolic activity. CONCLUSION: We identified mediators likely contributing to the disease-perpetuating mesenchymal-to-epithelial crosstalk in ILD. In our alveolar organoid co-cultures, we were able to highlight the importance of fibroblast-initiated aberrant epithelial differentiation and confirmed IL11 as a key player in fibrotic-ILD pathogenesis by unbiased fibroblast secretome analysis. AU - Kastlmeier, M.T. AU - Gonzalez-Rodriguez, E. AU - Cabanis , P. AU - Günther, E. AU - König, A.-C. AU - Han, L. AU - Hauck, S.M. AU - See, F. AU - Asgharpour, S. AU - Bukas, C. AU - Burgstaller, G. AU - Piraud, M. AU - Lehmann, M. AU - Hatz, R.A.* AU - Behr, J.* AU - Stöger, T. AU - Hilgendorff, A. AU - Voss, C. C1 - 67859 C2 - 54337 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Cytokine signaling converging on IL11 in ILD fibroblasts provokes aberrant epithelial differentiation signatures. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - Injuries to our skin trigger a cascade of spatially- and temporally-synchronized healing processes. During such endogenous wound repair, the role of fibroblasts is multifaceted, ranging from the activation and recruitment of innate immune cells through the synthesis and deposition of scar tissue to the conveyor belt-like transport of fascial connective tissue into wounds. A comprehensive understanding of fibroblast diversity and versatility in the healing machinery may help to decipher wound pathologies whilst laying the foundation for novel treatment modalities. In this review, we portray the diversity of fibroblasts and delineate their unique wound healing functions. In addition, we discuss future directions through a clinical-translational lens. AU - Knoedler, S. AU - Broichhausen, S.* AU - Guo, R. AU - Dai, R. AU - Knoedler, L.* AU - Kauke-Navarro, M.* AU - Diatta, F.* AU - Pomahac, B.* AU - Machens, H.G.* AU - Jiang, D. AU - Rinkevich, Y. C1 - 68343 C2 - 54762 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Fibroblasts – the cellular choreographers of wound healing. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - BACKGROUND: Kidney transplant recipients (KTRs) are at high risk for a severe course of coronavirus disease 2019 (COVID-19); thus, effective vaccination is critical. However, the achievement of protective immunogenicity is hampered by immunosuppressive therapies. We assessed cellular and humoral immunity and breakthrough infection rates in KTRs vaccinated with homologous and heterologous COVID-19 vaccination regimens. METHOD: We performed a comparative in-depth analysis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T-cell responses using multiplex Fluorospot assays and SARS-CoV-2-specific neutralizing antibodies (NAbs) between three-times homologously (n = 18) and heterologously (n = 8) vaccinated KTRs. RESULTS: We detected SARS-CoV-2-reactive T cells in 100% of KTRs upon third vaccination, with comparable frequencies, T-cell expression profiles, and relative interferon γ and interleukin 2 production per single cell between homologously and heterologously vaccinated KTRs. SARS-CoV-2-specific NAb positivity rates were significantly higher in heterologously (87.5%) compared to homologously vaccinated (50.0%) KTRs (P < 0.0001), whereas the magnitudes of NAb titers were comparable between both subcohorts after third vaccination. SARS-CoV-2 breakthrough infections occurred in equal numbers in homologously (38.9%) and heterologously (37.5%) vaccinated KTRs with mild-to-moderate courses of COVID-19. CONCLUSION: Our data support a more comprehensive assessment of not only humoral but also cellular SARS-CoV-2-specific immunity in KTRs to provide an in-depth understanding about the COVID-19 vaccine-induced immune response in a transplant setting. AU - Körber, N. AU - Holzmann-Littig, C.* AU - Wilkens, G. AU - Liao, B.H.* AU - Werz, M.L.* AU - Platen, L.* AU - Cheng, C.C.* AU - Tellenbach, M.* AU - Kappler, V.* AU - Lehner, V.* AU - Mijočević, H.* AU - Christa, C.* AU - Assfalg, V.* AU - Heemann, U.* AU - Schmaderer, C.* AU - Protzer, U. AU - Braunisch, M.C.* AU - Bauer, T. AU - Renders, L.* C1 - 67601 C2 - 53908 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Comparable cellular and humoral immunity upon homologous and heterologous COVID-19 vaccination regimens in kidney transplant recipients. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - Crosstalk between innate and adaptive immunity is pivotal for an efficient immune response and to maintain immune homeostasis under steady state conditions. As part of the innate immune system, type 2 innate lymphoid cells (ILC2s) have emerged as new important regulators of tissue homeostasis and repair by fine-tuning innate-adaptive immune cell crosstalk. ILC2s mediate either pro- or anti-inflammatory immune responses in a context dependent manner. Inflammation has proven to be a key driver of atherosclerosis, resembling the key underlying pathophysiology of cardiovascular disease (CVD). Notably, numerous studies point towards an atheroprotective role of ILC2s e.g., by mediating secretion of type-II cytokines (IL-5, IL-13, IL-9). Boosting these protective responses may be suitable for promising future therapy, although these protective cues are currently incompletely understood. Additionally, little is known about the mechanisms by which chemokine/chemokine receptor signaling shapes ILC2 functions in vascular inflammation and atherosclerosis. Hence, this review will focus on the latest findings regarding the protective and chemokine/chemokine receptor guided interplay between ILC2s and other immune cells like T and B cells, dendritic cells and macrophages in atherosclerosis. Further, we will elaborate on potential therapeutic implications which result or could be distilled from the dialogue of ILC2s with cells of the immune system in cardiovascular diseases. AU - Kral, M.* AU - van der Vorst, E.P.C.* AU - Surnov, A. AU - Weber, C.* AU - Döring, Y.* C1 - 68995 C2 - 53801 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - ILC2-mediated immune crosstalk in chronic (vascular) inflammation. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Premature ovarian failure (POF) is a major cause of infertility among women of reproductive age. Unfortunately, there is no effective treatment available currently. Researchers have shown that immune disorders play a significant role in the development of POF. Moreover, growing evidence suggest that Chitosan Oligosaccharides (COS), which act as critical immunomodulators, may have a key role in preventing and treating a range of immune related reproductive diseases. METHODS: KM mice (6-8 weeks) received a single intraperitoneal injection of cyclophosphamide (CY, 120mg/kg) and busulfan (BUS, 30mg/kg) to establish POF model. After completing the COS pre-treatment or post-treatment procedures, peritoneal resident macrophages (PRMs) were collected for neutral erythrophagocytosis assay to detect phagocytic activity. The thymus, spleen and ovary tissues were collected and weighed to calculate the organ indexes. Hematoxylin-eosin (HE) staining was performed to observe the histopathologic structure of those organs. The serum levels of estrogen (E2) and progesterone (P) were measured via the enzyme-linked immunosorbent assay (ELISA). The expression levels of immune factors including interleukin 2 (IL-2), interleukin 4 (IL-4), and tumor necrosis factor α (TNF-α), as well as germ cell markers Mouse Vasa Homologue (MVH) and Fragilis in ovarian tissue, were analyzed by Western blotting and qRT-PCR. In addition, ovarian cell senescence via p53/p21/p16 signaling was also detected. RESULTS: The phagocytic function of PRMs and the structural integrity of thymus and spleen were preserved by COS treatment. The levels of certain immune factors in the ovaries of CY/BUS- induced POF mice were found to be altered, manifested as IL-2 and TNF-α experiencing a significant decline, and IL-4 presenting a notable increase. Both pre-treatment and post-treatment with COS were shown to be protective effects against the damage to ovarian structure caused by CY/BUS. Senescence-associated β-galactosidase (SA-β-Gal) staining results showed that COS prevents CY/BUS-induced ovarian cell senescence. Additionally, COS regulated estrogen and progesterone levels, enhanced follicular development, and blocked ovarian cellular p53/p21/p16 signaling which participating in cell senescence. CONCLUSION: COS is a potent preventative and therapeutic medicine for premature ovarian failure by enhancing both the ovarian local and systemic immune response as well as inhibiting germ cell senescence. AU - Li, X.* AU - Ye, H. AU - Su, T.* AU - Hu, C.* AU - Huang, Y.* AU - Fu, X.* AU - Zhong, Z.* AU - Du, X.* AU - Zheng, Y.* C1 - 67895 C2 - 54373 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Immunity and reproduction protective effects of Chitosan Oligosaccharides in Cyclophosphamide/Busulfan-induced premature ovarian failure model mice. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - Allergic inflammation of the airways such as allergic asthma is a major health problem with growing incidence world-wide. One cardinal feature in severe type 2-dominated airway inflammation is the release of lipid mediators of the eicosanoid family that can either promote or dampen allergic inflammation. Macrophages are key producers of prostaglandins and leukotrienes which play diverse roles in allergic airway inflammation and thus require tight control. Using RNA- and ATAC-sequencing, liquid chromatography coupled to mass spectrometry (LC-MS/MS), enzyme immunoassays (EIA), gene expression analysis and in vivo models, we show that the aryl hydrocarbon receptor (AhR) contributes to this control via transcriptional regulation of lipid mediator synthesis enzymes in bone marrow-derived as well as in primary alveolar macrophages. In the absence or inhibition of AhR activity, multiple genes of both the prostaglandin and the leukotriene pathway were downregulated, resulting in lower synthesis of prostanoids, such as prostaglandin E2 (PGE2), and cysteinyl leukotrienes, e.g., Leukotriene C4 (LTC4). These AhR-dependent genes include PTGS1 encoding for the enzyme cyclooxygenase 1 (COX1) and ALOX5 encoding for the arachidonate 5-lipoxygenase (5-LO) both of which major upstream regulators of the prostanoid and leukotriene pathway, respectively. This regulation is independent of the activation stimulus and partially also detectable in unstimulated macrophages suggesting an important role of basal AhR activity for eicosanoid production in steady state macrophages. Lastly, we demonstrate that AhR deficiency in hematopoietic but not epithelial cells aggravates house dust mite induced allergic airway inflammation. These results suggest an essential role for AhR-dependent eicosanoid regulation in macrophages during homeostasis and inflammation. AU - Maier, A.-M. AU - Huth, K. AU - Alessandrini, F. AU - Schnautz, B. AU - Arifovic, A. AU - Riols, F. AU - Haid, M. AU - Koegler, A.* AU - Sameith, K.* AU - Schmidt-Weber, C.B. AU - Esser-von Bieren, J. AU - Ohnmacht, C. C1 - 67681 C2 - 53988 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - The aryl hydrocarbon receptor regulates lipid mediator production in alveolar macrophages. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - In the published article, there was an error in the author list, and author Fiona Henkel was erroneously excluded. The corrected author list appears below. Ann-Marie Maier1, Karsten Huth1, Francesca Alessandrini1, Fiona Henkel1, Benjamin Schnautz1, Anela Arifovic1, Fabien Riols2, Mark Haid2, Anja Koegler3, Katrin Sameith3, Carsten B Schmidt-Weber1,4, Julia Esser-von-Bieren1,5, Caspar Ohnmacht1 Fiona Henkel is therefore removed from the Acknowledgments. The corrected Acknowledgments appears below. We thank Johanna Grosch for help with the preparation of histology slides. New author Fiona Henkel is now part of the Author contributions. The corrected Author contributions statement appears below. A-MM performed most experiments and data analysis. KH, FA, BS and AA contributed to experiments and analysis. FH, FR and MH performed LC-MS/MS measurements. AK and KS performed bioinformatic analysis. CS-W and JE-B co-supervised the study. CO supervised the study, performed data interpretation, acquired funding and wrote the manuscript with input from all co-authors. All authors contributed to the article and approved the submitted version. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated. AU - Maier, A.-M. AU - Huth, K. AU - Alessandrini, F. AU - Henkel, F. AU - Schnautz, B. AU - Arifovic, A. AU - Riols, F. AU - Haid, M. AU - Koegler, A.* AU - Sameith, K.* AU - Schmidt-Weber, C.B. AU - Esser-von Bieren, J. AU - Ohnmacht, C. C1 - 68499 C2 - 54672 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Corrigendum: The aryl hydrocarbon receptor regulates lipid mediator production in alveolar macrophages(Front. Immunol., (2023), 14, (1157373), 10.3389/fimmu.2023.1157373). JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Patients with primary adrenal insufficiency (PAI) suffer from increased risk of infection, adrenal crises and have a higher mortality rate. Such dismal outcomes have been inferred to immune cell dysregulation because of unphysiological cortisol replacement. As the immune landscape of patients with different types of PAI has not been systematically explored, we set out to immunophenotype PAI patients with different causes of glucocorticoid (GC) deficiency. METHODS: This cross-sectional single center study includes 28 patients with congenital adrenal hyperplasia (CAH), 27 after bilateral adrenalectomy due to Cushing's syndrome (BADx), 21 with Addison's disease (AD) and 52 healthy controls. All patients with PAI were on a stable GC replacement regimen with a median dose of 25 mg hydrocortisone per day. Peripheral blood mononuclear cells were isolated from heparinized blood samples. Immune cell subsets were analyzed using multicolor flow cytometry after four-hour stimulation with phorbol myristate acetate and ionomycin. Natural killer (NK-) cell cytotoxicity and clock gene expression were investigated. RESULTS: The percentage of T helper cell subsets was downregulated in AD patients (Th1 p = 0.0024, Th2 p = 0.0157, Th17 p < 0.0001) compared to controls. Cytotoxic T cell subsets were reduced in AD (Tc1 p = 0.0075, Tc2 p = 0.0154) and CAH patients (Tc1 p = 0.0055, Tc2 p = 0.0012) compared to controls. NKCC was reduced in all subsets of PAI patients, with smallest changes in CAH. Degranulation marker CD107a expression was upregulated in BADx and AD, not in CAH patients compared to controls (BADx p < 0.0001; AD p = 0.0002). In contrast to NK cell activating receptors, NK cell inhibiting receptor CD94 was upregulated in BADx and AD, but not in CAH patients (p < 0.0001). Although modulation in clock gene expression could be confirmed in our patient subgroups, major interindividual-intergroup dissimilarities were not detected. DISCUSSION: In patients with different etiologies of PAI, distinct differences in T and NK cell-phenotypes became apparent despite the use of same GC preparation and dose. Our results highlight unsuspected differences in immune cell composition and function in PAI patients of different causes and suggest disease-specific alterations that might necessitate disease-specific treatment. AU - Nowotny, H.F.* AU - Marchant Seiter, T.* AU - Ju, J.* AU - Gottschlich, A.* AU - Schneider, H.* AU - Zopp, S.* AU - Vogel, F.* AU - Tschaidse, L.* AU - Auer, M.K.* AU - Lottspeich, C.* AU - Kobold, S. AU - Rothenfußer, S. AU - Beuschlein, F.* AU - Reincke, M.* AU - Braun, L.* AU - Reisch, N.* C1 - 68878 C2 - 53739 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Major immunophenotypic abnormalities in patients with primary adrenal insufficiency of different etiology. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - MALT1 is a core component of the CARD11-BCL10-MALT1 (CBM) signalosome, in which it acts as a scaffold and a protease to bridge T cell receptor (TCR) ligation to immune activation. As a scaffold, MALT1 binds to TRAF6, and T cell-specific TRAF6 ablation or destruction of MALT1-TRAF6 interaction provokes activation of conventional T (Tconv) effector cells. In contrast, MALT1 protease activity controls the development and suppressive function of regulatory T (Treg) cells in a T cell-intrinsic manner. Thus, complete loss of TRAF6 or selective inactivation of MALT1 catalytic function in mice skews the immune system towards autoimmune inflammation, but distinct mechanisms are responsible for these immune disorders. Here we demonstrate that TRAF6 deletion or MALT1 paracaspase inactivation are highly interdependent in causing the distinct immune pathologies. We crossed mice with T cell-specific TRAF6 ablation (Traf6-ΔT) and mice with a mutation rendering the MALT1 paracaspase dead in T cells (Malt1 PD-T) to yield Traf6-ΔT;Malt1 PD-T double mutant mice. These mice reveal that the autoimmune inflammation caused by TRAF6-ablation relies strictly on the function of the MALT1 protease to drive the activation of Tconv cells. Vice versa, despite the complete loss of Treg cells in Traf6-ΔT;Malt1 PD-T double mutant mice, inactivation of the MALT1 protease is unable to cause autoinflammation, because the Tconv effector cells are not activated in the absence of TRAF6. Consequentially, combined MALT1 paracaspase inactivation and TRAF6 deficiency in T cells mirrors the immunodeficiency seen upon T cell-specific MALT1 ablation. AU - O’Neill, T.J.* AU - Gewies, A. AU - Seeholzer, T. AU - Krappmann, D. C1 - 67418 C2 - 54141 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - TRAF6 controls T cell homeostasis by maintaining the equilibrium of MALT1 scaffolding and protease functions. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - GAD-alum given into lymph nodes to Type 1 diabetes (T1D) patients participating in a multicenter, randomized, placebo-controlled double-blind study seemed to have a positive effect for patients with DR3DQ2 haplotype, who showed better preservation of C-peptide than the placebo group. Here we compared the immunomodulatory effect of GAD-alum administered into lymph nodes of patients with T1D versus placebo with focus on patients with DR3DQ2 haplotype. Methods: GAD autoantibodies, GADA subclasses, GAD65-induced cytokine secretion (Luminex panel) and proliferation of peripheral mononuclear cells were analyzed in T1D patients (n=109) who received either three intra-lymphatic injections (one month apart) with 4 µg GAD-alum and oral vitamin D supplementation (2000 IE daily for 120 days), or placebo. Results: Higher GADA, GADA subclasses, GAD65-induced proliferation and cytokine secretion was observed in actively treated patients after the second injection of GAD-alum compared to the placebo group. Following the second injection of GAD-alum, actively treated subjects with DR3DQ2 haplotype had higher GAD65-induced secretion of several cytokine (IL4, IL5, IL7, IL10, IL13, IFNγ, GM-CSF and MIP1β) and proliferation compared to treated individuals without DR3DQ2. Stratification of samples from GAD-alum treated patients according to C-peptide preservation at 15 months revealed that “good responder” individuals with better preservation of C-peptide secretion, independently of the HLA haplotype, had increased GAD65-induced proliferation and IL13 secretion at 3 months, and a 2,5-fold increase of IL5 and IL10 as compared to “poor responders”. The second dose of GAD-alum also induced a more pronounced cytokine secretion in “good responders” with DR3DQ2, compared to few “good responders” without DR3DQ2 haplotype. Conclusion: Patients with DR3DQ2 haplotype had a distinct early cellular immune response to GAD-alum injections into the lymph node, and predominant GAD65-induced IL13 secretion and proliferation that seems to be associated with a better clinical outcome. If confirmed in the ongoing larger randomized double-blind placebo-controlled clinical trial (DIAGNODE-3), including only patients carrying DR3DQ2 haplotype, these results might be used as early surrogate markers for clinical efficacy. AU - Puente-Marin, S.* AU - Dietrich, F.* AU - Achenbach, P. AU - Barcenilla, H.* AU - Ludvigsson, J.* AU - Casas, R.* C1 - 67511 C2 - 54103 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Intralymphatic glutamic acid decarboxylase administration in type 1 diabetes patients induced a distinctive early immune response in patients with DR3DQ2 haplotype. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - Common flow cytometry-based methods used for functional assessment of antigen-specific T cells rely on de novo expression of intracellular cytokines or cell surface activation induced markers. They come with some limitations such as complex experimental setting, loss of cell viability and often high unspecific background which impairs assay sensitivity. We have previously shown that staining of activated ß2-integrins either with multimers of their ligand ICAM-1 or with a monoclonal antibody can serve as a functional marker detectable on T cells after minutes (CD8+) or few hours (CD4+) of activation. Here, we present a simple method for detection of activated ß2-integrins in combination with established cell surface activation induced markers. We observed that activated ß2-integrins were still detectable after 14 hours of stimulation, allowing their detection together with CD137 and CD154. Combinatorial gating of cells expressing activated ß2-integrins and CD137 or CD154 reduced background in unstimulated samples, increasing the signal-to-noise ratio and allowing improved assessment of low-frequency T cell responses. Extracellular staining of these markers highly correlated with production of intracellular cytokines IL-2, TNF or IFNγ in CD4+ and CD8+ T cells. As an exemplary application, SARS-CoV-2 spike-specific T cell responses were assessed in individuals after COVID-19 vaccination. This method should be useful for epitope discovery projects and for the simultaneous monitoring of low-frequency antigen-specific CD4+ and CD8+ T cell responses in various physiological situations. AU - Schöllhorn, A.* AU - Maia, A.* AU - Kimmerle, F.* AU - Born, J. AU - Rammensee, H.G.* AU - Dimitrov, S.* AU - Gouttefangeas, C.* C1 - 67402 C2 - 54158 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Staining of activated ß2-integrins in combination with CD137 and CD154 for sensitive identification of functional antigen-specific CD4+ and CD8+ T cells. JO - Front. Immunol. VL - 13 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - Background: Bladder urothelial carcinoma (BLCA) is associated with high mortality and recurrence. Although mRNA-based vaccines are promising treatment strategies for combating multiple solid cancers, their efficacy against BLCA remains unclear. We aimed to identify potential effective antigens of BLCA for the development of mRNA-based vaccines and screen for immune clusters to select appropriate candidates for vaccination. Methods: Gene expression microarray data and clinical information were retrieved from The Cancer Genome Atlas and GSE32894, respectively. The mRNA splicing patterns were obtained from the SpliceSeq portal. The cBioPortal for Cancer Genomics was used to visualize genetic alteration profiles. Furthermore, nonsense-mediated mRNA decay (NMD) analysis, correlation analysis, consensus clustering analysis, immune cell infiltration analysis, and weighted co-expression network analysis were conducted. Results: Six upregulated and mutated tumor antigens related to NMD, and infiltration of APCs were identified in patients with BLCA, including HP1BP3, OSBPL9, SSH3, ZCCHC8, FANCI, and EIF4A2. The patients were subdivided into two immune clusters (IC1 and IC2) with distinct clinical, cellular and molecular features. Patients in IC1 represented immunologically ‘hot’ phenotypes, whereas those in IC2 represented immunologically ‘cold’ phenotypes. Moreover, the survival rate was better in IC2 than in IC1, and the immune landscape of BLCA indicated significant inter-patient heterogeneity. Finally, CALD1, TGFB3, and ANXA6 were identified as key genes of BLCA through WGCNA analysis, and their mRNA expression levels were measured using qRT-PCR. Conclusion: HP1BP3, OSBPL9, SSH3, ZCCHC8, FANCI, and EIF4A2 were identified as potential antigens for developing mRNA-based vaccines against BLCA, and patients in IC2 might benefit more from vaccination. AU - Sun, Z.* AU - Jing, C. AU - Zhan, H.* AU - Guo, X.* AU - Suo, N.* AU - Kong, F.* AU - Tao, W.* AU - Xiao, C.* AU - Hu, D.* AU - Wang, H.* AU - Jiang, S.* C1 - 67427 C2 - 54142 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Identification of tumor antigens and immune landscapes for bladder urothelial carcinoma mRNA vaccine. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - INTRODUCTION: Inflammation is a key driver of morbidity in the vulnerable preterm infant exposed to pre- and postnatal hazards and significantly contributes to chronic lung disease, i.e. bronchopulmonary dysplasia (BPD). However, the early changes in innate immunity associated with BPD development are incompletely understood. METHODS: In very immature preterm infants below 32 weeks gestational age (GA; n=30 infants), monocyte subtypes were identified by Flow Cytometry at birth and throughout the postnatal course including intracellular TNF expression upon LPS stimulation. Complementing these measurements, cytokine end growth factor expression profiles (Luminex® xMAP®; n=110 infants) as well as gene expression profiles (CodeLinkTM Human I Bioarray; n=22) were characterized at birth. RESULTS: The abundance of monocyte subtypes differed between preterm and term neonates at birth. Specifically, CD14++CD16+ (intermediate) monocytes demonstrated a dependency on PMA and elevated levels of nonclassical (CD14+CD16++) monocytes characterized preterm infants with developing BPD. Postnatally, lung injury was associated with an increase in intermediate monocytes, while high levels of nonclassical monocytes persisted. Both subtypes were revealed as the main source of intracellular TNF-α expression in the preterm infant. We identified a cytokine and growth factor expression profile in cord blood specimen of preterm infants with developing BPD that corresponded to the disease-dependent regulation of monocyte abundances. Multivariate modeling of protein profiles revealed FGF2, sIL-2 Rα, MCP-1, MIP1a, and TNF-α as predictors of BPD when considering GA. Transcriptome analysis demonstrated genes predicting BPD to be overrepresented in inflammatory pathways with increased disease severity characterized by the regulation of immune and defense response pathways and upstream regulator analysis confirmed TNF-α, interleukin (IL) -6, and interferon α as the highest activated cytokines in more severe disease. Whereas all BPD cases showed downstream activation of chemotaxis and activation of inflammatory response pathways, more severe cases were characterized by an additional activation of reactive oxygen species (ROS) synthesis. DISCUSSION: In the present study, we identified the early postnatal presence of nonclassical (CD14+CD16++) and intermediate (CD14++CD16+) monocytes as a critical characteristic of BPD development including a specific response pattern of monocyte subtypes to lung injury. Pathophysiological insight was provided by the protein and transcriptome signature identified at birth, centered around monocyte and corresponding granulocyte activation and highlighting TNFα as a critical regulator in infants with developing BPD. The disease severity-dependent expression patterns could inform future diagnostic and treatment strategies targeting the monocytic cell and its progeny. AU - Windhorst, A.C.* AU - Heydarian, M. AU - Schwarz, M. AU - Oak, P. AU - Forster, K.* AU - Frankenberger, M. AU - Gonzalez-Rodriguez, E. AU - Zhang, X. AU - Ehrhardt, H.* AU - Hübener, C.* AU - Flemmer, A.W.* AU - Hossain, H.* AU - Stöger, T. AU - Schulz, C.* AU - Hilgendorff, A. C1 - 67675 C2 - 53982 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Monocyte signature as a predictor of chronic lung disease in the preterm infant. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - [This corrects the article DOI: 10.3389/fimmu.2023.1112608.]. AU - Windhorst, A.C.* AU - Heydarian, M. AU - Schwarz, M. AU - Oak, P. AU - Forster, K.* AU - Frankenberger, M. AU - Gonzalez-Rodriguez, E. AU - Zhang, X. AU - Ehrhardt, H.* AU - Hübener, C.* AU - Flemmer, A.W.* AU - Hossain, H.* AU - Stöger, T. AU - Schulz, C.* AU - Hilgendorff, A. C1 - 67797 C2 - 54275 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Corrigendum: Monocyte signature as a predictor of chronic lung disease in the preterm infant. JO - Front. Immunol. VL - 14 PB - Frontiers Media Sa PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - Foxp3+ regulatory T (Treg) cells of thymic (tTreg) and peripheral (pTreg) developmental origin are thought to synergistically act to ensure immune homeostasis, with self-reactive tTreg cells primarily constraining autoimmune responses. Here we exploited a Foxp3-dependent reporter with thymus-specific GFP/Cre activity to selectively ablate either tTreg (ΔtTreg) or pTreg (ΔpTreg) cell development, while sparing the respective sister populations. We found that, in contrast to the tTreg cell behavior in ΔpTreg mice, pTreg cells acquired a highly activated suppressor phenotype and replenished the Treg cell pool of ΔtTreg mice on a non-autoimmune C57BL/6 background. Despite the absence of tTreg cells, pTreg cells prevented early mortality and fatal autoimmunity commonly observed in Foxp3-deficient models of complete Treg cell deficiency, and largely maintained immune tolerance even as the ΔtTreg mice aged. However, only two generations of backcrossing to the autoimmune-prone non-obese diabetic (NOD) background were sufficient to cause severe disease lethality associated with different, partially overlapping patterns of organ-specific autoimmunity. This included a particularly severe form of autoimmune diabetes characterized by an early onset and abrogation of the sex bias usually observed in the NOD mouse model of human type 1 diabetes. Genetic association studies further allowed us to define a small set of autoimmune risk loci sufficient to promote β cell autoimmunity, including genes known to impinge on Treg cell biology. Overall, these studies show an unexpectedly high functional adaptability of pTreg cells, emphasizing their important role as mediators of bystander effects to ensure self-tolerance. AU - Yilmazer, A.* AU - Zevla, D.M.* AU - Malmkvist, R.* AU - Rodríguez, C.A.B.* AU - Undurraga, P.* AU - Kirgin, E.* AU - Boernert, M.* AU - Voehringer, D.* AU - Kershaw, O.* AU - Schlenner, S.* AU - Kretschmer, K. C1 - 69019 C2 - 53804 TI - Selective ablation of thymic and peripheral Foxp3+ regulatory T cell development. JO - Front. Immunol. VL - 14 PY - 2023 SN - 1664-3224 ER - TY - JOUR AB - The lung epithelial barrier serves as a guardian towards environmental insults and responds to allergen encounter with a cascade of immune reactions that can possibly lead to inflammation. Whether the environmental sensor aryl hydrocarbon receptor (AhR) together with its downstream targets cytochrome P450 (CYP1) family members contribute to the regulation of allergic airway inflammation remains unexplored. By employing knockout mice for AhR and for single CYP1 family members, we found that AhR-/- and CYP1B1-/- but not CYP1A1-/- or CYP1A2-/- animals display enhanced allergic airway inflammation compared to WT. Expression analysis, immunofluorescence staining of murine and human lung sections and bone marrow chimeras suggest an important role of CYP1B1 in non-hematopoietic lung epithelial cells to prevent exacerbation of allergic airway inflammation. Transcriptional analysis of murine and human lung epithelial cells indicates a functional link of AhR to barrier protection/inflammatory mediator signaling upon allergen challenge. In contrast, CYP1B1 deficiency leads to enhanced expression and activity of CYP1A1 in lung epithelial cells and to an increased availability of the AhR ligand kynurenic acid following allergen challenge. Thus, differential CYP1 family member expression and signaling via the AhR in epithelial cells represents an immunoregulatory layer protecting the lung from exacerbation of allergic airway inflammation. AU - Alessandrini, F. AU - de Jong, R.J. AU - Wimmer, M. AU - Maier, A.-M. AU - Fernandez, I.E. AU - Hils, M.* AU - Buters, J.T.M. AU - Biedermann, T. AU - Zissler, U.M. AU - Hoffmann, C. AU - Esser-von Bieren, J. AU - Schmidt-Weber, C.B. AU - Ohnmacht, C. C1 - 65541 C2 - 52730 TI - Lung epithelial CYP1 activity regulates aryl hydrocarbon receptor dependent allergic airway inflammation. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Despite its high prevalence, the cellular and molecular mechanisms of chronic obstructive pulmonary disease (COPD) are far from being understood. Here, we determine disease-related changes in cellular and molecular compositions within the alveolar space and peripheral blood of a cohort of COPD patients and controls. Myeloid cells were the largest cellular compartment in the alveolar space with invading monocytes and proliferating macrophages elevated in COPD. Modeling cell-to-cell communication, signaling pathway usage, and transcription factor binding predicts TGF-β1 to be a major upstream regulator of transcriptional changes in alveolar macrophages of COPD patients. Functionally, macrophages in COPD showed reduced antigen presentation capacity, accumulation of cholesteryl ester, reduced cellular chemotaxis, and mitochondrial dysfunction, reminiscent of impaired immune activation. AU - Bassler, K.* AU - Fujii, W.* AU - Kapellos, T.S.* AU - Dudkin, E.* AU - Reusch, N.* AU - Horne, A.* AU - Reiz, B.* AU - Luecken, M. AU - Osei-Sarpong, C.* AU - Warnat-Herresthal, S.* AU - Bonaguro, L.* AU - Schulte-Schrepping, J.* AU - Wagner, A.* AU - Guenther, P.* AU - Pizarro, C.* AU - Schreiber, T.* AU - Knöll, R.* AU - Holsten, L.* AU - Kroeger, C.* AU - De Domenico, E.* AU - Becker, M.* AU - Haendler, K.* AU - Wohnhaas, C.T.* AU - Baumgartner, F.* AU - Koehler, M.* AU - Theis, H.* AU - Kraut, M.* AU - Wadsworth, M.H.* AU - Hughes, T.K.* AU - Ferreira, H.J.* AU - Hinkley, E.* AU - Kaltheuner, I.H.* AU - Geyer, M.* AU - Thiele, C.* AU - Shalek, A.K.* AU - Feisst, A.* AU - Thomas, D.* AU - Dickten, H.* AU - Beyer, M.* AU - Baum, P.* AU - Yosef, N.* AU - Aschenbrenner, A.C.* AU - Ulas, T.* AU - Hasenauer, J. AU - Theis, F.J. AU - Skowasch, D.* AU - Schultze, J.L.* C1 - 65933 C2 - 52563 TI - Alveolar macrophages in early stage COPD show functional deviations with properties of impaired immune activation. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Post-transcriptional gene regulation by RNA-binding proteins (RBPs) is important in the prevention of inflammatory and autoimmune diseases. With respect to T cell activation and differentiation, the RBPs Roquin-1/2 and Regnase-1 play pivotal roles by inducing degradation and/or translational silencing of target mRNAs. These targets encode important proinflammatory mediators and thus Roquin and Regnase-1 functions dampen cellular programs that can lead to inflammation and autoimmune disease. Recent findings demonstrate direct physical interaction of both RBPs. Here, we propose that cooperativity of trans-acting factors may be more generally used to reinforce the regulatory impact on selected targets and promote specific cell fate decisions. We develop this concept for Roquin and Regnase-1 function in resting and activated T cells and discuss the involvement in autoimmunity as well as how the therapeutic potential can be used in anti-tumor therapies. AU - Behrens, G.* AU - Heissmeyer, V. C1 - 64511 C2 - 52242 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Cooperation of RNA-binding proteins - a focus on roquin function in T cells. JO - Front. Immunol. VL - 13 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - The immune system is a complex and sophisticated biological system, spanning multiple levels of complexity, from the molecular level to that of tissue. Our current understanding of its function and complexity, of the heterogeneity of leukocytes, is a result of decades of concentrated efforts to delineate cellular markers using conventional methods of antibody screening and antigen identification. In mammalian models, this led to in-depth understanding of individual leukocyte subsets, their phenotypes, and their roles in health and disease. The field was further propelled forward by the development of single-cell (sc) RNA-seq technologies, offering an even broader and more integrated view of how cells work together to generate a particular response. Consequently, the adoption of scRNA-seq revealed the unexpected plasticity and heterogeneity of leukocyte populations and shifted several long-standing paradigms of immunology. This review article highlights the unprecedented opportunities offered by scRNA-seq technology to unveil the individual contributions of leukocyte subsets and their crosstalk in generating the overall immune responses in bony fishes. Single-cell transcriptomics allow identifying unseen relationships, and formulating novel hypotheses tailored for teleost species, without the need to rely on the limited number of fish-specific antibodies and pre-selected markers. Several recent studies on single-cell transcriptomes of fish have already identified previously unnoticed expression signatures and provided astonishing insights into the diversity of teleost leukocytes and the evolution of vertebrate immunity. Without a doubt, scRNA-seq in tandem with bioinformatics tools and state-of-the-art methods, will facilitate studying the teleost immune system by not only defining key markers, but also teaching us about lymphoid tissue organization, development/differentiation, cell-cell interactions, antigen receptor repertoires, states of health and disease, all across time and space in fishes. These advances will invite more researchers to develop the tools necessary to explore the immunology of fishes, which remain non-conventional animal models from which we have much to learn. AU - Chan, J.T.H.* AU - Kadri, S. AU - Köllner, B.* AU - Rebl, A.* AU - Korytář, T.* C1 - 64317 C2 - 52182 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - RNA-seq of single fish cells – seeking out the leukocytes mediating immunity in teleost fishes. JO - Front. Immunol. VL - 13 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Despite the availability of an effective prophylactic vaccine, 820,000 people die annually of hepatitis B virus (HBV)-related liver disease according to WHO. Since current antiviral therapies do not provide a curative treatment for the 296 million HBV carriers around the globe, novel strategies to cure HBV are urgently needed. A promising approach is the redirection of T cells towards HBV-infected hepatocytes employing chimeric antigen receptors or T-cell engager antibodies. We recently described the effective redirection of T cells employing a second-generation chimeric antigen receptor directed against the envelope protein of hepatitis B virus on the surface of infected cells (S-CAR) as well as bispecific antibodies that engage CD3 or CD28 on T cells employing the identical HBV envelope protein (HBVenv) binder. In this study, we added a trispecific antibody comprising all three moieties to the tool-box. Cytotoxic and non-cytolytic antiviral activities of these bi- and trispecific T-cell engager antibodies were assessed in co-cultures of human PBMC with HBV-positive hepatoma cells, and compared to that of S-CAR-grafted T cells. Activation of T cells via the S-CAR or by either a combination of the CD3- and CD28-targeting bispecific antibodies or the trispecific antibody allowed for specific elimination of HBV-positive target cells. While S-CAR-grafted effector T cells displayed faster killing kinetics, combinatory treatment with the bispecific antibodies or single treatment with the trispecific antibody was associated with a more pronounced cytokine release. Clearance of viral antigens and elimination of the HBV persistence form, the covalently closed circular (ccc) DNA, through cytolytic as well as cytokine-mediated activity was observed in all three settings with the combination of bispecific antibodies showing the strongest non-cytolytic, cytokine-mediated antiviral effect. Taken together, we demonstrate that bi- and trispecific T-cell engager antibodies can serve as a potent, off-the-shelf alternative to S-CAR-grafted T cells to cure HBV. AU - Debelec-Butuner, B. AU - Quitt, O. AU - Schreiber, S. AU - Momburg, F.* AU - Wisskirchen, K. AU - Protzer, U. C1 - 66752 C2 - 53288 TI - Activation of distinct antiviral T-cell immunity: A comparison of bi- and trispecific T-cell engager antibodies with a chimeric antigen receptor targeting HBV envelope proteins. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Heterozygous TREX1 mutations are associated with monogenic familial chilblain lupus and represent a risk factor for developing systemic lupus erythematosus. These interferonopathies originate from chronic type I interferon stimulation due to sensing of inadequately accumulating nucleic acids. We here analysed the composition of dendritic cell (DC) subsets, central stimulators of immune responses, in patients with TREX1 deficiency. We performed single-cell RNA-sequencing of peripheral blood DCs and monocytes from two patients with familial chilblain lupus and heterozygous mutations in TREX1 and from controls. Type I interferon pathway genes were strongly upregulated in patients. Cell frequencies of the myeloid and plasmacytoid DC and of monocyte populations in patients and controls were similar, but we describe a novel DC subpopulation highly enriched in patients: a myeloid DC CD1C+ subpopulation characterized by the expression of LMNA, EMP1 and a type I interferon- stimulated gene profile. The presence of this defined subpopulation was confirmed in a second cohort of patients and controls by flow cytometry, also revealing that an increased percentage of patient’s cells in the subcluster express costimulatory molecules. We identified a novel type I interferon responsive myeloid DC subpopulation, that might be important for the perpetuation of TREX1-induced chilblain lupus and other type I interferonopathies. AU - Eugster, A.* AU - Müller, D.* AU - Gompf, A.* AU - Reinhardt, S.* AU - Lindner, A.* AU - Ashton, M.P.* AU - Zimmermann, N.* AU - Beissert, S.* AU - Bonifacio, E. AU - Günther, C.* C1 - 65821 C2 - 52939 TI - A novel type I interferon primed dendritic cell subpopulation in TREX1 mutant chilblain lupus patients. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - [This corrects the article DOI: 10.3389/fimmu.2022.897500.]. AU - Eugster, A.* AU - Müller, D.* AU - Gompf, A.* AU - Reinhardt, S.* AU - Lindner, A.* AU - Ashton, M.P.* AU - Zimmermann, N.* AU - Beissert, S.* AU - Bonifacio, E. AU - Günther, C.* C1 - 66984 C2 - 53423 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Corrigendum: A novel type I interferon primed dendritic cell subpopulation in TREX1 mutant chilblain lupus patients. JO - Front. Immunol. VL - 13 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Inflammatory bowel disease (IBD) is a multifactorial disorder triggered by imbalances of the microbiome and immune dysregulations in genetically susceptible individuals. Several mouse and human studies have demonstrated that multimeric inflammasomes are critical regulators of host defense and gut homeostasis by modulating immune responses to pathogen- or damage-associated molecular patterns. In the context of IBD, excessive production of pro-inflammatory Interleukin-1β has been detected in patient-derived intestinal tissues and correlated with the disease severity or failure to respond to anti-tumor necrosis factor therapy. Correspondingly, genome-wide association studies have suggested that single nucleotide polymorphisms in inflammasome components might be associated with risk of IBD development. The relevance of inflammasomes in controlling human intestinal homeostasis has been further exemplified by the discovery of very early onset IBD (VEO-IBD) patients with monogenic defects affecting different molecules in the complex regulatory network of inflammasome activity. This review provides an overview of known causative monogenic entities of VEO-IBD associated with altered inflammasome activity. A better understanding of the molecular mechanisms controlling inflammasomes in monogenic VEO-IBD may open novel therapeutic avenues for rare and common inflammatory diseases. AU - Illig, D.* AU - Kotlarz, D.M. C1 - 66979 C2 - 53393 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Dysregulated inflammasome activity in intestinal inflammation - Insights from patients with very early onset IBD. JO - Front. Immunol. VL - 13 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - The alpha-Gal epitope (α-Gal) with the determining element galactose-α1,3-galactose can lead to clinically relevant allergic reactions and rejections in xenotransplantation. These immune reactions can develop because humans are devoid of this carbohydrate due to evolutionary loss of the enzyme α1,3-galactosyltransferase (GGTA1). In addition, up to 1% of human IgG antibodies are directed against α-Gal, but the stimulus for the induction of anti-α-Gal antibodies is still unclear. Commensal bacteria have been suggested as a causal factor for this induction as α-Gal binding tools such as lectins were found to stain cultivated bacteria isolated from the intestinal tract. Currently available tools for the detection of the definite α-Gal epitope, however, are cross-reactive, or have limited affinity and, hence, offer restricted possibilities for application. In this study, we describe a novel monoclonal IgG1 antibody (27H8) specific for the α-Gal epitope. The 27H8 antibody was generated by immunization of Ggta1 knockout mice and displays a high affinity towards synthetic and naturally occurring α-Gal in various applications. Using this novel tool, we found that intestinal bacteria reported to be α-Gal positive cannot be stained with 27H8 questioning whether commensal bacteria express the native α-Gal epitope at all. AU - Kreft, L. AU - Schepers, A. AU - Hils, M.* AU - Swiontek, K.* AU - Flatley, A. AU - Janowski, R. AU - Mirzaei, M.K. AU - Dittmar, M. AU - Chakrapani, N.* AU - Desai, M.S.* AU - Eyerich, S. AU - Deng, L. AU - Niessing, D. AU - Fischer, K.* AU - Feederle, R. AU - Blank, S. AU - Schmidt-Weber, C.B. AU - Hilger, C.* AU - Biedermann, T.* AU - Ohnmacht, C. C1 - 66001 C2 - 53033 TI - A novel monoclonal IgG1 antibody specific for Galactose-alpha-1,3-galactose questions alpha-Gal epitope expression by bacteria. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Activation of CD40-signaling contributes to the initiation, progression and drug resistance of B cell lymphomas. We contributed to this knowledge by showing that constitutive CD40-signaling in B cells induces B cell hyperplasia and finally B cell lymphoma development in transgenic mice. CD40 activates, among others, the non-canonical NF-ĸB signaling, which is constitutively activated in several human B cell lymphomas and is therefore presumed to contribute to lymphopathogenesis. This prompted us to study the regulatory role of the non-canonical NF-ĸB transcription factor RelB in lymphomagenesis. To this end, we crossed mice expressing a constitutively active CD40 receptor in B cells with conditional RelB-KO mice. Ablation of RelB attenuated pre-malignant B cell expansion, and resulted in an impaired survival and activation of long-term CD40-stimulated B cells. Furthermore, we found that hyperactivation of non-canonical NF-кB signaling enhances the retention of B cells in the follicles of secondary lymphoid organs. RNA-Seq-analysis revealed that several genes involved in B-cell migration, survival, proliferation and cytokine signaling govern the transcriptional differences modulated by the ablation of RelB in long-term CD40-stimulated B cells. Inactivation of RelB did not abrogate lymphoma development. However, lymphomas occurred with a lower incidence and had a longer latency period. In summary, our data suggest that RelB, although it is not strictly required for malignant transformation, accelerates the lymphomagenesis of long-term CD40-stimulated B cells by regulating genes involved in migration, survival and cytokine signaling. AU - Kuhn, L. AU - Valentin, S. AU - Stojanovic, K. AU - Strobl, D.C. AU - Babushku, T. AU - Wang, Y. AU - Rambold, U. AU - Scheffler, L. AU - Grath, S.* AU - John-Robbert, D.* AU - Blum, H.* AU - Feuchtinger, A. AU - Blutke, A. AU - Weih, F.* AU - Kitamura, D.* AU - Rad, R.* AU - Strobl, L.J. AU - Zimber-Strobl, U. C1 - 66233 C2 - 52963 TI - RelB contributes to the survival, migration and lymphomagenesis of B cells with constitutively active CD40 signaling. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AU - Meng, Z.* AU - Liu, J.* AU - Kosinska, A. AU - Lu, M.* C1 - 64735 C2 - 52429 TI - Editorial: Targeting the immune system to treat hepatitis B virus infection. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - TGF-β1 is known to have a pro-inflammatory impact by inducing Th9 and Th17 cells, while it also induces anti-inflammatory Treg cells (Tregs). In the context of allergic airway inflammation (AAI) its dual role can be of critical importance in influencing the outcome of the disease. Here we demonstrate that TGF-β is a major player in AAI by driving effector T cells, while Tregs differentiate independently. Induction of experimental AAI and airway hyperreactivity in a mouse model with inducible genetic ablation of the gene encoding for TGFβ-receptor 2 (Tgfbr2) on CD4+T cells significantly reduced the disease phenotype. Further, it blocked the induction of pro-inflammatory T cell frequencies (Th2, Th9, Th17), but increased Treg cells. To translate these findings into a human clinically relevant context, Th2, Th9 and Treg cells were quantified both locally in induced sputum and systemically in blood of allergic rhinitis and asthma patients with or without allergen-specific immunotherapy (AIT). Natural allergen exposure induced local and systemic Th2, Th9, and reduced Tregs cells, while therapeutic allergen exposure by AIT suppressed Th2 and Th9 cell frequencies along with TGF-β and IL-9 secretion. Altogether, these findings support that neutralization of TGF-β represents a viable therapeutic option in allergy and asthma, not posing the risk of immune dysregulation by impacting Tregs cells. AU - Musiol, S. AU - Alessandrini, F. AU - Jakwerth, C.A. AU - Chaker, A. AU - Schneider, E. AU - Guerth, F. AU - Schnautz, B. AU - Grosch, J. AU - Ghiordanescu, I. AU - Ullmann, J.T. AU - Kau, J. AU - Plaschke, M. AU - Haak, S.* AU - Buch, T.* AU - Schmidt-Weber, C.B. AU - Zissler, U.M. C1 - 64150 C2 - 52092 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - TGF-β1 drives inflammatory Th cell but not treg cell compartment upon allergen exposure. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Understanding immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial to contain the COVID-19 pandemic. Using a multiplex approach, serum IgG responses against the whole SARS-CoV-2 proteome and the nucleocapsid proteins of endemic human coronaviruses (HCoVs) were measured in SARS-CoV-2-infected donors and healthy controls. COVID-19 severity strongly correlated with IgG responses against the nucleocapsid (N) of SARS-CoV-2 and possibly with the number of viral antigens targeted. Furthermore, a strong correlation between COVID-19 severity and serum responses against N of endemic alpha- but not betacoronaviruses was detected. This correlation was neither caused by cross-reactivity of antibodies, nor by a general boosting effect of SARS-CoV-2 infection on pre-existing humoral immunity. These findings raise the prospect of a potential disease progression marker for COVID-19 severity that allows for early stratification of infected individuals. AU - Nückel, J.* AU - Planatscher, E.* AU - Mohr, A.W.* AU - Deichl, K.* AU - Mijočević, H. AU - Feuerherd, M. AU - Wolff, L.S. AU - Erber, J.* AU - Schneider, J.* AU - Quante, M.* AU - Winter, C.* AU - Ruland, J.* AU - Hapfelmeier, A.* AU - Hammerschmidt, W. AU - Moosmann, A.* AU - Protzer, U. AU - Behrends, U.* AU - Mautner, J. C1 - 66264 C2 - 53124 TI - Association between IgG responses against the nucleocapsid proteins of alphacoronaviruses and COVID-19 severity. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Quantum dots (QDs), are one kind of nanoscale semiconductor crystals with specific electronic and optical properties, offering near-infrared mission and chemically active surfaces. Increasing interest for QDs exists in developing theranostics platforms for bioapplications such as imaging, drug delivery and therapy. Here we summarized QDs' biomedical applications, toxicity, and immunological effects on the respiratory system. Bioapplications of QDs in lung include biomedical imaging, drug delivery, bio-sensing or diagnosis and therapy. Generically, toxic effects of nanoparticles are related to the generation of oxidative stresses with subsequent DNA damage and decreased lung cells viability in vitro and in vivo because of release of toxic metal ions or the features of QDs like its surface charge. Lastly, pulmonary immunological effects of QDs mainly include proinflammatory cytokines release and recruiting innate leukocytes or adaptive T cells. AU - Ren, L.* AU - Wang, L.* AU - Rehberg, M. AU - Stöger, T. AU - Zhang, J.* AU - Chen, S.* C1 - 64157 C2 - 52095 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Applications and immunological effects of quantum dots on respiratory system. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Kidney renal clear cell carcinoma (KIRC) is one of the most prevalent primary malignancies with high heterogeneity in the urological system. Growing evidence implies that lactate is a significant carbon source for cell metabolism and plays a vital role in tumor development, maintenance, and therapeutic response. However, the global influence of lactate-related genes (LRGs) on prognostic significance, tumor microenvironment characteristics, and therapeutic response has not been comprehensively elucidated in patients with KIRC. In the present study, we collected RNA sequencing and clinical data of KIRC from The Cancer Genome Atlas (TCGA), E-MTAB-1980, and GSE22541 cohorts. Unsupervised clustering of 17 differentially expressed LRG profiles divided the samples into three clusters with distinct immune characteristics. Three genes (FBP1, HADH, and TYMP) were then identified to construct a lactate-related prognostic signature (LRPS) using the least absolute shrinkage and selection operator (LASSO) and Cox regression analyses. The novel signature exhibited excellent robustness and predictive ability for the overall survival of patients. In addition, the constructed nomogram based on the LRPS-based risk scores and clinical factors (age, gender, tumor grade, and stage) showed a robust predictive performance. Furthermore, patients classified by risk scores had distinguishable immune status, tumor mutation burden, response to immunotherapy, and sensitivity to drugs. In conclusion, we developed an LRPS for KIRC that was closely related to the immune landscape and therapeutic response. This LRPS may guide clinicians to make more precise and personalized treatment decisions for KIRC patients. AU - Sun, Z.J.* AU - Tao, W.* AU - Guo, X.* AU - Jing, C. AU - Zhang, M.* AU - Wang, Z.* AU - Kong, F.* AU - Suo, N.* AU - Jiang, S.* AU - Wang, H.* C1 - 64512 C2 - 52244 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Construction of a lactate-related prognostic signature for predicting prognosis, tumor microenvironment, and immune response in kidney renal clear cell carcinoma. JO - Front. Immunol. VL - 13 PB - Frontiers Media Sa PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Background and aims: There is growing interest in T cell-based immune therapies for a functional cure of chronic HBV infection including check-point inhibition, T cell-targeted vaccines or TCR-grafted effector cells. All these approaches depend on recognition of HLA class I-presented viral peptides. The HBV core region 18-27 is an immunodominant target of CD8+ T cells and represents the prime target for T cell-based therapies. Here, a high-resolution analysis of the core18-27 specific CD8+ T cell and the selected escape pathways was performed. Methods: HLA class I typing and viral sequence analyses were performed for 464 patients with chronic HBV infection. HBV-specific CD8+ T-cell responses against the prototype and epitope variants were characterized by flow cytometry. Results: Consistent with promiscuous presentation of the core18-27 epitope, antigen-specific T cells were detected in patients carrying HLA-A*02:01, HLA-B*35:01, HLA-B*35:03 or HLA-B*51:01. Sequence analysis confirmed reproducible selection pressure on the core18-27 epitope in the context of these alleles. Interestingly, the selected immune escape pathways depend on the presenting HLA-class I-molecule. Although cross-reactive T cells were observed, some epitope variants achieved functional escape by impaired TCR-interaction or disturbed antigen processing. Of note, selection of epitope variants was exclusively observed in HBeAg negative HBV infection and here, detection of variants associated with significantly greater magnitude of the CD8 T cell response compared to absence of variants. Conclusion: The core18-27 epitope is highly variable and under heavy selection pressure in the context of different HLA class I-molecules. Some epitope variants showed evidence for impaired antigen processing and reduced presentation. Viruses carrying such escape substitutions will be less susceptible to CD8+ T cell responses and should be considered for T cell-based therapy strategies. AU - Walker, A.* AU - Schwarz, T.* AU - Brinkmann-Paulukat, J.* AU - Wisskirchen, K. AU - Menne, C.* AU - Alizei, E.S.* AU - Kefalakes, H.* AU - Theissen, M.* AU - Hoffmann, D.* AU - Schulze zur Wiesch, J.* AU - Maini, M.K.* AU - Cornberg, M.* AU - Kraft, A.R.M.* AU - Keitel, V.* AU - Bock, H.H.* AU - Horn, P.A.* AU - Thimme, R.* AU - Wedemeyer, H.* AU - Heinemann, F.M.* AU - Luedde, T.* AU - Neumann-Haefelin, C.* AU - Protzer, U. AU - Timm, J.* C1 - 66829 C2 - 53304 TI - Immune escape pathways from the HBV core18-27 CD8 T cell response are driven by individual HLA class I alleles. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - Skin malignant melanoma is a highly aggressive skin tumor, which is also a major cause of skin cancer-related mortality. It can spread from a relatively small primary tumor and metastasize to multiple locations, including lymph nodes, lungs, liver, bone, and brain. What’s more metastatic melanoma is the main cause of its high mortality. Among all organs, the lung is one of the most common distant metastatic sites of melanoma, and the mortality rate of melanoma lung metastasis is also very high. Elucidating the mechanisms involved in the pulmonary metastasis of cutaneous melanoma will not only help to provide possible explanations for its etiology and progression but may also help to provide potential new therapeutic targets for its treatment. Increasing evidence suggests that tumor-associated macrophages (TAMs) play an important regulatory role in the migration and metastasis of various malignant tumors. Tumor-targeted therapy, targeting tumor-associated macrophages is thus attracting attention, particularly for advanced tumors and metastatic tumors. However, the relevant role of tumor-associated macrophages in cutaneous melanoma lung metastasis is still unclear. This review will present an overview of the origin, classification, polarization, recruitment, regulation and targeting treatment of tumor-associated macrophages, as well as the soluble mediators involved in these processes and a summary of their possible role in lung metastasis from cutaneous malignant melanoma. This review particularly aims to provide insight into mechanisms and potential therapeutic targets to readers, interested in pulmonary metastasis melanoma. AU - Xiong, K.* AU - Qi, M.* AU - Stöger, T. AU - Zhang, J.* AU - Chen, S.* C1 - 66254 C2 - 53120 TI - The role of tumor-associated macrophages and soluble mediators in pulmonary metastatic melanoma. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AB - The cellular events that dictate the initiation of the complement pathway in ocular degeneration, such as age-related macular degeneration (AMD), is poorly understood. Using gene expression analysis (single cell and bulk), mass spectrometry, and immunohistochemistry, we dissected the role of multiple retinal and choroidal cell types in determining the complement homeostasis. Our scRNA-seq data show that the cellular response to early AMD is more robust in the choroid, particularly in fibroblasts, pericytes and endothelial cells. In late AMD, complement changes were more prominent in the retina especially with the expression of the classical pathway initiators. Notably, we found a spatial preference for these differences. Overall, this study provides insights into the heterogeneity of cellular responses for complement expression and the cooperation of neighboring cells to complete the pathway in healthy and AMD eyes. Further, our findings provide new cellular targets for therapies directed at complement. AU - Zauhar, R.* AU - Biber, J.* AU - Jabri, Y.* AU - Kim, M.* AU - Hu, J.* AU - Kaplan, L.* AU - Pfaller, A.M.* AU - Schaefer, N.* AU - Enzmann, V.* AU - Schloetzer-Schrehardt, U.* AU - Straub, T.* AU - Hauck, S.M. AU - Gamlin, P.D.* AU - McFerrin, M.B.* AU - Messinger, J.* AU - Strang, C.E.* AU - Curcio, C.A.* AU - Dana, N.* AU - Pauly, D.* AU - Grosche, A.* AU - Li, M.* AU - Stambolian, D.* C1 - 65598 C2 - 52379 TI - As in real estate, location matters: Cellular expression of complement varies between macular and peripheral regions of the retina and supporting Tissues. JO - Front. Immunol. VL - 13 PY - 2022 SN - 1664-3224 ER - TY - JOUR AU - Blank, S. AU - Hilger, C.* C1 - 61919 C2 - 50337 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Editorial: Novel advances in allergy diagnosis and treatment. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Background: Adaptive immune responses to structural proteins of the virion play a crucial role in protection against coronavirus disease 2019 (COVID-19). We therefore studied T cell responses against multiple SARS-CoV-2 structural proteins in a large cohort using a simple, fast, and high-throughput approach. Methods: An automated interferon gamma release assay (IGRA) for the Nucleocapsid (NC)-, Membrane (M)-, Spike-C-terminus (SCT)-, and N-terminus-protein (SNT)-specific T cell responses was performed using fresh whole blood from study subjects with convalescent, confirmed COVID-19 (n = 177, more than 200 days post infection), exposed household members (n = 145), and unexposed controls (n = 85). SARS-CoV-2-specific antibodies were assessed using Elecsys® Anti-SARS-CoV-2 (Ro-N-Ig) and Anti-SARS-CoV-2-ELISA (IgG) (EI-S1-IgG). Results: 156 of 177 (88%) previously PCR confirmed cases were still positive by Ro-N-Ig more than 200 days after infection. In T cells, most frequently the M-protein was targeted by 88% seropositive, PCR confirmed cases, followed by SCT (85%), NC (82%), and SNT (73%), whereas each of these antigens was recognized by less than 14% of non-exposed control subjects. Broad targeting of these structural virion proteins was characteristic of convalescent SARS-CoV-2 infection; 68% of all seropositive individuals targeted all four tested antigens. Indeed, anti-NC antibody titer correlated loosely, but significantly with the magnitude and breadth of the SARS-CoV-2-specific T cell response. Age, sex, and body mass index were comparable between the different groups. Conclusion: SARS-CoV-2 seropositivity correlates with broad T cell reactivity of the structural virus proteins at 200 days after infection and beyond. The SARS-CoV-2-IGRA can facilitate large scale determination of SARS-CoV-2-specific T cell responses with high accuracy against multiple targets. AU - Brand, I.* AU - Gilberg, L.* AU - Bruger, J.* AU - Garí, M. AU - Wieser, A.* AU - Eser, T.M.* AU - Frese, J.* AU - Ahmed, M.I.M.* AU - Rubio-Acero, R.* AU - Guggenbüehl Noller, J.M.* AU - Castelletti, N.* AU - Diekmannshemke, J. AU - Thiesbrummel, S. AU - Huynh, D.* AU - Winter, S.* AU - Kroidl, I.* AU - Fuchs, C. AU - Hoelscher, M.* AU - Roider, J.* AU - Kobold, S. AU - Pritsch, M.* AU - Geldmacher, C.* C1 - 62185 C2 - 50686 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Broad T cell targeting of structural proteins after SARS-CoV-2 infection: High throughput assessment of T cell reactivity using an automated interferon gamma release assay. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - COVID-19, the disease caused by SARS-CoV-2 infection, can assume a highly variable disease course, ranging from asymptomatic infection, which constitutes the majority of cases, to severe respiratory failure. This implies a diverse host immune response to SARS-CoV-2. However, the immunological underpinnings underlying these divergent disease courses remain elusive. We therefore set out to longitudinally characterize immune signatures of convalescent COVID-19 patients stratified according to their disease severity. Our unique convalescent COVID-19 cohort consists of 74 patients not confounded by comorbidities. This is the first study of which we are aware that excludes immune abrogations associated with non-SARS-CoV-2 related risk factors of disease severity. Patients were followed up and analyzed longitudinally (2, 4 and 6 weeks after infection) by high-dimensional flow cytometric profiling of peripheral blood mononuclear cells (PBMCs), in-depth serum analytics, and transcriptomics. Immune phenotypes were correlated to disease severity. Convalescence was overall associated with uniform immune signatures, but distinct immune signatures for mildly versus severely affected patients were detectable within a 2-week time window after infection. AU - Chu, C.F.* AU - Sabath, F.* AU - Fibi-Smetana, S.* AU - Sun, S.* AU - Öllinger, R.* AU - Nößner, E. AU - Chao, Y.Y.* AU - Rinke, L.L.* AU - Winheim, E.* AU - Rad, R.* AU - Krug, A.B.* AU - Taher, L.* AU - Zielinski, C.E.* C1 - 62986 C2 - 51180 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Convalescent COVID-19 patients without comorbidities display similar immunophenotypes over time despite divergent disease severities. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Whole genome sequencing of Epstein-Barr virus (EBV) isolates from around the world has uncovered pervasive strain heterogeneity, but the forces driving strain diversification and the impact on immune recognition remained largely unknown. Using a data mining approach, we analyzed more than 300 T-cell epitopes in 168 published EBV strains. Polymorphisms were detected in approximately 65% of all CD8+ and 80% of all CD4+ T-cell epitopes and these numbers further increased when epitope flanking regions were included. Polymorphisms in CD8+ T-cell epitopes often involved MHC anchor residues and resulted in changes of the amino acid subgroup, suggesting that only a limited number of conserved T-cell epitopes may represent generic target antigens against different viral strains. Although considered the prototypic EBV strain, the rather low degree of overlap with most other viral strains implied that B95.8 may not represent the ideal reference strain for T-cell epitopes. Instead, a combinatorial library of consensus epitopes may provide better targets for diagnostic and therapeutic purposes when the infecting strain is unknown. Polymorphisms were significantly enriched in epitope versus non-epitope protein sequences, implicating immune selection in driving strain diversification. Remarkably, CD4+ T-cell epitopes in EBNA2, EBNA-LP, and the EBNA3 family appeared to be under negative selection pressure, hinting towards a beneficial role of immune responses against these latency type III antigens in virus biology. These findings validate this immunoinformatics approach for providing novel insight into immune targets and the intricate relationship of host defense and virus evolution that may also pertain to other pathogens. AU - Cirac, A.* AU - Poirey, R.* AU - Dieckmeyer, M.* AU - Witter, K.* AU - Delecluse, H.J.* AU - Behrends, U. AU - Mautner, J. C1 - 63934 C2 - 51706 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Immunoinformatic analysis reveals antigenic heterogeneity of Epstein-Barr virus is immune-driven. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Background: Treatment of B-cell malignancies with CD19-directed chimeric antigen receptor (CAR) T-cells marked a new era in immunotherapy, which yet has to be successfully adopted to solid cancers. Epigenetic inhibitors of DNA methyltransferases (DNMTi) and histone deacetylases (HDACi) can induce broad changes in gene expression of malignant cells, thus making these inhibitors interesting combination partners for immunotherapeutic approaches. Methods: Urothelial carcinoma cell lines (UCC) and benign uroepithelial HBLAK cells pretreated with the DNMTi decitabine or the HDACi romidepsin were co-incubated with CAR T-cells directed against EGFR or CD44v6, and subsequent cytotoxicity assays were performed. Effects on T-cell cytotoxicity and surface antigen expression on UCC were determined by flow cytometry. We also performed next-generation mRNA sequencing of inhibitor-treated UCC and siRNA-mediated knockdown of potential regulators of CAR T-cell killing. Results: Exposure to decitabine but not romidepsin enhanced CAR T-cell cytotoxicity towards all UCC lines, but not towards the benign HBLAK cells. Increased killing could neither be attributed to enhanced target antigen expression (EGFR and CD44v6) nor fully explained by changes in the T-cell ligands PD-L1, PD-L2, ICAM-1, or CD95. Instead, gene expression analysis suggested that regulators of cell survival and apoptosis were differentially induced by the treatment. Decitabine altered the balance between survival and apoptosis factors towards an apoptosis-sensitive state associated with increased CAR T-cell killing, while romidepsin, at least partially, tilted this balance in the opposite direction. Knockdown experiments with siRNA in UCC confirmed BID and BCL2L1/BCLX as two key factors for the altered susceptibility of the UCC. Conclusion: Our data suggest that the combination of decitabine with CAR T-cell therapy is an attractive novel therapeutic approach to enhance tumor-specific killing of bladder cancer. Since BID and BCL2L1 are essential determinants for the susceptibility of a wide variety of malignant cells, their targeting might be additionally suitable for combination with immunotherapies, e.g., CAR T-cells or checkpoint inhibitors in other malignancies. AU - Grunewald, C.M.* AU - Haist, C.* AU - König, C.* AU - Petzsch, P.* AU - Bister, A.* AU - Nößner, E. AU - Wiek, C.* AU - Scheckenbach, K.* AU - Köhrer, K.* AU - Niegisch, G.* AU - Hanenberg, H.* AU - Hoffmann, M.J.* C1 - 63715 C2 - 51674 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Epigenetic priming of bladder cancer cells with decitabine increases cytotoxicity of human EGFR and CD44v6 CAR engineered T-cells. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - The human Vγ9Vδ2 T cell is a unique cell type that holds great potential in immunotherapy of cancer. In particular, the therapeutic potential of this cell type in adoptive cell therapy (ACT) has gained interest. In this regard optimization of in vitro expansion methods and functional characterization is desirable. We show that Vγ9Vδ2 T cells, expanded in vitro with zoledronic acid (Zometa or ZOL) and Interleukin-2 (IL-2), are efficient cancer cell killers with a trend towards increased killing efficacy after prolonged expansion time. Thus, Vγ9Vδ2 T cells expanded for 25 days in vitro killed prostate cancer cells more efficiently than Vγ9Vδ2 T cells expanded for 9 days. These data are supported by phenotype characteristics, showing increased expression of CD56 and NKG2D over time, reaching above 90% positive cells after 25 days of expansion. At the early stage of expansion, we demonstrate that Vγ9Vδ2 T cells are capable of cross-presenting tumor antigens. In this regard, our data show that Vγ9Vδ2 T cells can take up tumor-associated antigens (TAA) gp100, MART-1 and MAGE-A3 - either as long peptide or recombinant protein - and then present TAA-derived peptides on the cell surface in the context of HLA class I molecules, demonstrated by their recognition as targets by peptide-specific CD8 T cells. Importantly, we show that cross-presentation is impaired by the proteasome inhibitor lactacystin. In conclusion, our data indicate that Vγ9Vδ2 T cells are broadly tumor-specific killers with the additional ability to cross-present MHC class I-restricted peptides, thereby inducing or supporting tumor-specific αβTCR CD8 T cell responses. The dual functionality is dynamic during in vitro expansion, yet, both functions are of interest to explore in ACT for cancer therapy. AU - Holmen Olofsson, G.* AU - Idorn, M.* AU - Carnaz Simões, A.M.* AU - Aehnlich, P.* AU - Skadborg, S.K.* AU - Nößner, E. AU - Debets, R.* AU - Moser, B.* AU - Met, Ö.* AU - Thor Straten, P.* C1 - 62318 C2 - 50770 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Vγ9Vδ2 T cells concurrently kill cancer cells and cross-present tumor antigens. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - A higher diversity of food items introduced in the first year of life has been inversely related to subsequent development of asthma. In the current analysis, we applied latent class analysis (LCA) to systematically assess feeding patterns and to relate them to asthma risk at school age. PASTURE (N=1133) and LUKAS2 (N=228) are prospective birth cohort studies designed to evaluate protective and risk factors for atopic diseases, including dietary patterns. Feeding practices were reported by parents in monthly diaries between the 4th and 12th month of life. For 17 common food items parents indicated frequency of feeding during the last 4 weeks in 4 categories. The resulting 153 ordinal variables were entered in a LCA. The intestinal microbiome was assessed at the age of 12 months by 16S rRNA sequencing. Data on feeding practice with at least one reported time point was available in 1042 of the 1133 recruited children. Best LCA model fit was achieved by the 4-class solution. One class showed an elevated risk of asthma at age 6 as compared to the other classes (adjusted odds ratio (aOR): 8.47, 95% CI 2.52-28.56, p = 0.001) and was characterized by daily meat consumption and rare consumption of milk and yoghurt. A refined LCA restricted to meat, milk, and yoghurt confirmed the asthma risk effect of a particular class in PASTURE and independently in LUKAS2, which we thus termed unbalanced meat consumption (UMC). The effect of UMC was particularly strong for non-atopic asthma and asthma irrespectively of early bronchitis (aOR: 17.0, 95% CI 5.2-56.1, p < 0.001). UMC fostered growth of iron scavenging bacteria such as Acinetobacter (aOR: 1.28, 95% CI 1.00-1.63, p = 0.048), which was also related to asthma (aOR: 1.55, 95% CI 1.18-2.03, p = 0.001). When reconstructing bacterial metabolic pathways from 16S rRNA sequencing data, biosynthesis of siderophore group nonribosomal peptides emerged as top hit (aOR: 1.58, 95% CI 1.13-2.19, p = 0.007). By a data-driven approach we found a pattern of overly meat consumption at the expense of other protein sources to confer risk of asthma. Microbiome analysis of fecal samples pointed towards overgrowth of iron-dependent bacteria and bacterial iron metabolism as a potential explanation. AU - Hose, A.J.* AU - Pagani, G. AU - Karvonen, A.M.* AU - Kirjavainen, P.V.* AU - Roduit, C.* AU - Genuneit, J.* AU - Schmaußer-Hechfellner, E. AU - Depner, M. AU - Frei, R.* AU - Lauener, R.* AU - Riedler, J.* AU - Schaub, B.* AU - Fuchs, O.* AU - von Mutius, E. AU - Divaret-Chauveau, A.* AU - Pekkanen, J.* AU - Ege, M.J.* C1 - 62025 C2 - 50596 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Excessive unbalanced meat consumption in the first year of life increases asthma risk in the PASTURE and LUKAS2 birth cohorts. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - T-cell therapy with T cells that are re-directed to hepatitis B virus (HBV)-infected cells by virus-specific receptors is a promising therapeutic approach for treatment of chronic hepatitis B and HBV-associated cancer. Due to the high number of target cells, however, side effects such as cytokine release syndrome or hepatotoxicity may limit safety. A safeguard mechanism, which allows depletion of transferred T cells on demand, would thus be an interesting means to increase confidence in this approach. In this study, T cells were generated by retroviral transduction to express either an HBV-specific chimeric antigen receptor (S-CAR) or T-cell receptor (TCR), and in addition either inducible caspase 9 (iC9) or herpes simplex virus thymidine kinase (HSV-TK) as a safety switch. Real-time cytotoxicity assays using HBV-replicating hepatoma cells as targets revealed that activation of both safety switches stopped cytotoxicity of S-CAR- or TCR-transduced T cells within less than one hour. In vivo, induction of iC9 led to a strong and rapid reduction of transferred S-CAR T cells adoptively transferred into AAV-HBV-infected immune incompetent mice. One to six hours after injection of the iC9 dimerizer, over 90% reduction of S-CAR T cells in the blood and the spleen and of over 99% in the liver was observed, thereby limiting hepatotoxicity and stopping cytokine secretion. Simultaneously, however, the antiviral effect of S-CAR T cells was diminished because remaining S-CAR T cells were mostly non-functional and could not be restimulated with HBsAg. A second induction of iC9 was only able to deplete T cells in the liver. In conclusion, T cells co-expressing iC9 and HBV-specific receptors efficiently recognize and kill HBV-replicating cells. Induction of T-cell death via iC9 proved to be an efficient means to deplete transferred T cells in vitro and in vivo containing unwanted hepatotoxicity. AU - Klopp, A. AU - Schreiber, S. AU - Kosinska, A. AU - Pulé, M.* AU - Protzer, U. AU - Wisskirchen, K. C1 - 63350 C2 - 51484 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Depletion of T cells via inducible caspase 9 increases safety of adoptive T-cell therapy against chronic hepatitis B. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Background: The cellular mechanisms involved in the lack of protective antibody response after hepatitis B vaccination are still rather unclear. Regulatory B cells (Breg) known as modulators of B-and T-cell responses may contribute to poor vaccine responsiveness. The current study aimed to investigate the role of regulatory B cells (Breg) in hepatitis B vaccine non-responsiveness after immunization with second- or third-generation hepatitis B vaccines. Method: We performed comparative phenotypic and frequency analysis of Breg subsets (CD24+CD27+ and CD24highCD38high Breg) in second-generation hepatitis B vaccine non-responders (2nd HBvac NR, n = 11) and responders (2nd HBvac R, n = 8) before (d0), on day 7 (d7), and 28 (d28) after booster vaccination. Cryopreserved peripheral blood mononuclear cells were stimulated ex vivo with a combination of CpG, PMA, and Ionomycin (CpG+P/I) and analyzed for numbers and IL-10 expression levels of Breg by flow cytometry-based analyses. Results: Flow cytometry-based analyses revealed elevated frequencies of CD24+CD27+ Breg at all time points and significantly higher frequencies of CD24highCD38high Breg on d0 (p = 0.004) and 28 (p = 0.012) in 2nd HBvac NR compared to 2nd HBvac R. In parallel, we observed significantly lower levels of CpG+P/I-induced IL-10 expression levels of CD24+CD27+ and CD24highCD38high Breg (d0: p < 0.0001; d7: p = 0.0004; d28: p = 0.0003 and d0: p = 0.016; d7: p = 0.016, respectively) in 2nd HBvac NR compared to 2nd HBvac R before and after booster immunization. Frequencies of CD24+CD27+ and CD24highCD38high Breg significantly decreased after third-generation hepatitis B booster vaccination (d7: p = 0.014; d28: p = 0.032 and d7: p = 0.045, respectively), whereas IL-10 expression levels of both Breg subsets remained stable. Conclusion: Here we report significantly higher frequencies of CD24highCD38high Breg in parallel with significantly lower IL-10 expression levels of CD24+CD27+ and CD24highCD38high Breg in 2nd HBvac NR compared to 2nd HBvac R. Anti-HBs seroconversion accompanied by a decrease of Breg numbers after booster immunization with a third-generation hepatitis B vaccine could indicate a positive effect of third-generation hepatitis B vaccines on Breg-mediated immunomodulation in hepatitis B vaccine non-responders. AU - Körber, N. AU - Pohl, L. AU - Weinberger, B.* AU - Grubeck-Loebenstein, B.* AU - Wawer, A.* AU - Knolle, P.A.* AU - Roggendorf, H.* AU - Protzer, U. AU - Bauer, T. C1 - 63107 C2 - 51313 TI - Hepatitis B vaccine non-responders show higher frequencies of CD24highCD38high regulatory B cells and lower levels of IL-10 expression compared to responders. JO - Front. Immunol. VL - 12 PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Replication competent vesicular stomatitis virus (VSV) is the basis of a vaccine against Ebola and VSV strains are developed as oncolytic viruses. Both functions depend on the ability of VSV to induce adequate amounts of interferon-α/β. It is therefore important to understand how VSV triggers interferon responses. VSV activates innate immunity via retinoic acid-inducible gene I (RIG-I), a sensor for viral RNA. Our results show that VSV needs to replicate for a robust interferon response. Analysis of RIG-I-associated RNA identified a copy-back defective-interfering (DI) genome and full-length viral genomes as main trigger of RIG-I. VSV stocks depleted of DI genomes lost most of their interferon-stimulating activity. The remaining full-length genome and leader-N-read-through sequences, however, still triggered RIG-I. Awareness for DI genomes as trigger of innate immune responses will help to standardize DI genome content and to purposefully deplete or use DI genomes as natural adjuvants in VSV-based therapeutics. AU - Linder, A.* AU - Bothe, V.* AU - Linder, N.* AU - Schwarzlmueller, P.* AU - Dahlström, F.* AU - Bartenhagen, C.* AU - Dugas, M.* AU - Pandey, D.* AU - Thorn-Seshold, J. AU - Boehmer, D.F.R.* AU - Koenig, L.M.* AU - Kobold, S.* AU - Schnurr, M.* AU - Raedler, J.* AU - Spielmann, G.* AU - Karimzadeh, H. AU - Schmidt, A.* AU - Endres, S. AU - Rothenfußer, S. C1 - 62108 C2 - 50654 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Defective interfering genomes and the full-length viral genome trigger RIG-I after infection with vesicular stomatitis virus in a replication dependent manner. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Increasing evidence suggests that post-translational peptide splicing can play a role in the immune response under pathological conditions. This seems to be particularly relevant in Type 1 Diabetes (T1D) since post-translationally spliced epitopes derived from T1D-associated antigens have been identified among those peptides bound to Human Leucocyte Antigen (HLA) class I and II complexes. Their immunogenicity has been confirmed through CD4 and CD8 T cell-mediated responses in T1D patients. Spliced peptides theoretically have a large sequence variability. This might increase the frequency of viral-human zwitter peptides, i.e. peptides that share a complete sequence homology irrespective of whether they originate from human or viral antigens, thereby impinging upon the discrimination between self and non-self antigens by T cells. This might increase the risk of autoimmune responses triggered by viral infections. Since enteroviruses and other viral infections have historically been associated with T1D, we investigated whether cis-spliced peptides derived from selected viruses might be able to trigger CD8 T cell-mediated autoimmunity. We computed in silico viral-human non-spliced and cis-spliced zwitter epitope candidates, and prioritized peptide candidates based on: (i) their binding affinity to HLA class I complexes, (ii) human pancreatic β cell and medullary thymic epithelial cell (mTEC) antigens’ mRNA expression, (iii) antigen association with T1D, and (iv) potential hotspot regions in those antigens. Neglecting potential T cell receptor (TCR) degeneracy, no viral-human zwitter non-spliced peptide was found to be an optimal candidate to trigger a virus-induced CD8 T cell response against human pancreatic β cells. Conversely, we identified some zwitter peptide candidates, which may be produced by proteasome-catalyzed peptide splicing, and might increase the likelihood of pancreatic β cells recognition by virus-specific CD8 T cell clones, therefore promoting β cell destruction in the context of viral infections. + + + + + AU - Mishto, M.* AU - Mansurkhodzhaev, A.* AU - Rodriguez-Calvo, T. AU - Liepe, J.* C1 - 61921 C2 - 50507 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Potential mimicry of viral and pancreatic β cell antigens through non-spliced and cis-spliced zwitter epitope candidates in type 1 diabetes. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Activation of co-stimulatory pathways in cytotoxic T lymphocytes expressing chimeric antigen receptors (CARs) have proven to boost effector activity, tumor rejection and long-term T cell persistence. When using antigen-specific T cell receptors (TCR) instead of CARs, the lack of co-stimulatory signals hampers robust antitumoral response, hence limiting clinical efficacy. In solid tumors, tumor stroma poses an additional hurdle through hindrance of infiltration and active inhibition. Our project aimed at generating chimeric co-stimulatory switch proteins (CSP) consisting of intracellular co-stimulatory domains (ICD) fused to extracellular protein domains (ECD) for which ligands are expressed in solid tumors. The ECD of CD40L was selected for combination with the ICD from the CD28 protein. With this approach, it was expected to not only provide co-stimulation and strengthen the TCR signaling, but also, through the CD40L ECD, facilitate the activation of tumor-resident antigen-presenting cells (APCs), modulate activation of tumor endothelium and induce TCR-MHC independent apoptotic effect on tumor cells. Since CD28 and CD40L belong to different classes of transmembrane proteins (type I and type II, respectively), creating a chimeric protein presented a structural and functional challenge. We present solutions to this challenge describing different CSP formats that were successfully expressed in human T cells along with an antigen-specific TCR. The level of surface expression of the CSPs depended on their distinct design and the state of T cell activation. In particular, CSPs were upregulated by TCR stimulation and downregulated following interaction with CD40 on target cells. Ligation of the CSP in the context of TCR-stimulation modulated intracellular signaling cascades and led to improved TCR-induced cytokine secretion and cytotoxicity. Moreover, the CD40L ECD exhibited activity as evidenced by effective maturation and activation of B cells and DCs. CD40L:CD28 CSPs are a new type of switch proteins designed to exert dual beneficial antitumor effect by acting directly on the gene-modified T cells and simultaneously on tumor cells and tumor-supporting cells of the TME. The observed effects suggest that they constitute a promising tool to be included in the engineering process of T cells to endow them with complementary features for improved performance in the tumor milieu. AU - Olguín-Contreras, L.F. AU - Mendler, A.N. AU - Popowicz, G.M. AU - Hu, B. AU - Nößner, E. C1 - 63846 C2 - 51555 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Double strike approach for tumor attack: Engineering T cells using a CD40L:CD28 chimeric co-stimulatory switch protein for enhanced tumor targeting in adoptive cell therapy. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - The mechanisms underlying type 1 diabetes (T1D) pathogenesis remain largely unknown. While autoantibodies to pancreatic beta-cell antigens are often the first biological response and thereby a useful biomarker for identifying individuals in early stages of T1D, their role in T1D pathogenesis is not well understood. Recognition of these antigenic targets by autoreactive T-cells plays a pathological role in T1D development. Recently, several beta-cell neoantigens have been described, indicating that both neoantigens and known T1D antigens escape central or peripheral tolerance. Several questions regarding the mechanisms by which tolerance is broken in T1D remain unanswered. Further delineating the timing and nature of antigenic responses could allow their use as biomarkers to improve staging, as targets for therapeutic intervention, and lead to a better understanding of the mechanisms leading to loss of tolerance. Multiple factors that contribute to cellular stress may result in the generation of beta-cell derived neoepitopes and contribute to autoimmunity. Understanding the cellular mechanisms that induce beta-cells to produce neoantigens has direct implications on development of therapies to intercept T1D disease progression. In this perspective, we will discuss evidence for the role of neoantigens in the pathogenesis of T1D, including antigenic responses and cellular mechanisms. We will additionally discuss the pathways leading to neoepitope formation and the cross talk between the immune system and the beta-cells in this regard. Ultimately, delineating the timing of neoepitope generation in T1D pathogenesis will determine their role as biomarkers as well as therapeutic targets. AU - Rodriguez-Calvo, T. AU - Johnson, J.D.* AU - Overbergh, L.* AU - Dunne, J.L.* C1 - 61933 C2 - 50520 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Neoepitopes in type 1 diabetes: Etiological insights, biomarkers and therapeutic targets. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - In human type 1 diabetes and animal models of the disease, a diverse assortment of immune cells infiltrates the pancreatic islets. CD8+ T cells are well represented within infiltrates and HLA multimer staining of pancreas sections provides clear evidence that islet epitope reactive T cells are present within autoimmune lesions. These bona fide effectors have been a key research focus because these cells represent an intellectually attractive culprit for β cell destruction. However, T cell receptors are highly diverse in human insulitis. This suggests correspondingly broad antigen specificity, which includes a majority of T cells for which there is no evidence of islet-specific reactivity. The presence of "non-cognate" T cells in insulitis raises suspicion that their role could be beyond that of an innocent bystander. In this perspective, we consider the potential pathogenic contribution of non-islet-reactive T cells. Our intellectual framework will be that of a criminal investigation. Having arraigned islet-specific CD8+ T cells for the murder of pancreatic β cells, we then turn our attention to the non-target immune cells present in human insulitis and consider the possible regulatory, benign, or effector roles that they may play in disease. Considering available evidence, we overview the case that can be made that non-islet-reactive infiltrating T cells should be suspected as co-conspirators or accessories to the crime and suggest some possible routes forward for reaching a better understanding of their role in disease. AU - Rodriguez-Calvo, T. AU - Christoffersson, G.* AU - Bender, C.* AU - Von Herrath, M.G.* AU - Mallone, R.* AU - Kent, S.C.* AU - James, E.A.* C1 - 62420 C2 - 50842 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Means, motive, and opportunity: Do non-islet-reactive infiltrating T cells contribute to autoimmunity in type 1 diabetes? JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - We have previously shown that conformational change in the β2-integrin is a very early activation marker that can be detected with fluorescent multimers of its ligand intercellular adhesion molecule (ICAM)-1 for rapid assessment of antigen-specific CD8+ T cells. In this study, we describe a modified protocol of this assay for sensitive detection of functional antigen-specific CD4+ T cells using a monoclonal antibody (clone m24 Ab) specific for the open, high-affinity conformation of the β2-integrin. The kinetics of β2-integrin activation was different on CD4+ and CD8+ T cells (several hours vs. few minutes, respectively); however, m24 Ab readily stained both cell types 4-6 h after antigen stimulation. With this protocol, we were able to monitor ex vivo effector and memory CD4+ and CD8+ T cells specific for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), cytomegalovirus (CMV), Epstein-Barr virus (EBV), and hepatitis B virus (HBV) in whole blood or cryopreserved peripheral blood mononuclear cells (PBMCs) of infected or vaccinated individuals. By costaining β2-integrin with m24 and CD154 Abs, we assessed extremely low frequencies of polyfunctional CD4+ T cell responses. The novel assay used in this study allows very sensitive and simultaneous screening of both CD4+ and CD8+ T cell reactivities, with versatile applicability in clinical and vaccination studies. AU - Schöllhorn, A.* AU - Schuhmacher, J.* AU - Besedovsky, L.* AU - Fendel, R.* AU - Jensen, A.T.R.* AU - Stevanović, S.* AU - Lange, T.* AU - Rammensee, H.G.* AU - Born, J. AU - Gouttefangeas, C.* AU - Dimitrov, S.I.* C1 - 61852 C2 - 50192 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Integrin activation enables sensitive detection of functional CD4+ and CD8+ T cells: Application to characterize SARS-CoV-2 immunity. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Immunodeficient mice engrafted with a functional human immune system [Human immune system (HIS) mice] have paved the way to major advances for personalized medicine and translation of immune-based therapies. One prerequisite for advancing personalized medicine is modeling the immune system of individuals or disease groups in a preclinical setting. HIS mice engrafted with peripheral blood mononuclear cells have provided fundamental insights in underlying mechanisms guiding immune activation vs. regulation in several diseases including cancer. However, the development of Graft-vs.-host disease restrains relevant long-term studies in HIS mice. Alternatively, engraftment with hematopoietic stem cells (HSCs) enables mimicking different disease stages, however, low frequencies of HSCs in peripheral blood of adults impede engraftment efficacy. One possibility to overcome those limitations is the use of patient-derived induced pluripotent stem cells (iPSCs) reprogrammed into HSCs, a challenging process which has recently seen major advances. Personalized HIS mice bridge research in mice and human diseases thereby facilitating the translation of immunomodulatory therapies. Regulatory T cells (Tregs) are important mediators of immune suppression and thereby contribute to tumor immune evasion, which has made them a central target for cancer immunotherapies. Importantly, studying Tregs in the human immune system in vivo in HIS mice will help to determine requirements for efficient Treg-targeting. In this review article, we discuss advances on personalized HIS models using reprogrammed iPSCs and review the use of HIS mice to study requirements for efficient targeting of human Tregs for personalized cancer immunotherapies. AU - Serr, I. AU - Kral, M. AU - Scherm, M.G. AU - Daniel, C. C1 - 61437 C2 - 50246 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Advances in human immune system mouse models for personalized treg-based immunotherapies. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Regulatory T cells (Tregs) are key mediators of peripheral self-tolerance and alterations in their frequencies, stability, and function have been linked to autoimmunity. The antigen-specific induction of Tregs is a long-envisioned goal for the treatment of autoimmune diseases given reduced side effects compared to general immunosuppressive therapies. However, the translation of antigen-specific Treg inducing therapies for the treatment or prevention of autoimmune diseases into the clinic remains challenging. In this mini review, we will discuss promising results for antigen-specific Treg therapies in allergy and specific challenges for such therapies in autoimmune diseases, with a focus on type 1 diabetes (T1D). We will furthermore discuss opportunities for antigen-specific Treg therapies in T1D, including combinatorial strategies and tissue-specific Treg targeting. Specifically, we will highlight recent advances in miRNA-targeting as a means to foster Tregs in autoimmunity. Additionally, we will discuss advances and perspectives of computational strategies for the detailed analysis of tissue-specific Tregs on the single-cell level. AU - Serr, I. AU - Drost, F.* AU - Schubert, B. AU - Daniel, C. C1 - 62745 C2 - 51041 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Antigen-specific Treg therapy in type 1 diabetes - challenges and opportunities. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AU - von Mutius, E. C1 - 61768 C2 - 50217 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - The “Hygiene Hypothesis” and the lessons learnt from farm studies. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Type 1 diabetes (T1D) represents a hallmark of the fatal multiorgan autoimmune syndrome affecting humans with abrogated Foxp3+ regulatory T (Treg) cell function due to Foxp3 gene mutations, but whether the loss of Foxp3+ Treg cell activity is indeed sufficient to promote β cell autoimmunity requires further scrutiny. As opposed to human Treg cell deficiency, β cell autoimmunity has not been observed in non-autoimmune-prone mice with constitutive Foxp3 deficiency or after diphtheria toxin receptor (DTR)-mediated ablation of Foxp3+ Treg cells. In the spontaneous nonobese diabetic (NOD) mouse model of T1D, constitutive Foxp3 deficiency did not result in invasive insulitis and hyperglycemia, and previous studies on Foxp3+ Treg cell ablation focused on Foxp3DTR NOD mice, in which expression of a transgenic BDC2.5 T cell receptor (TCR) restricted the CD4+ TCR repertoire to a single diabetogenic specificity. Here we revisited the effect of acute Foxp3+ Treg cell ablation on β cell autoimmunity in NOD mice in the context of a polyclonal TCR repertoire. For this, we took advantage of the well-established DTR/GFP transgene of DEREG mice, which allows for specific ablation of Foxp3+ Treg cells without promoting catastrophic autoimmune diseases. We show that the transient loss of Foxp3+ Treg cells in prediabetic NOD.DEREG mice is sufficient to precipitate severe insulitis and persistent hyperglycemia within 5 days after DT administration. Importantly, DT-treated NOD.DEREG mice preserved many clinical features of spontaneous diabetes progression in the NOD model, including a prominent role of diabetogenic CD8+ T cells in terminal β cell destruction. Despite the severity of destructive β cell autoimmunity, anti-CD3 mAb therapy of DT-treated mice interfered with the progression to overt diabetes, indicating that the novel NOD.DEREG model can be exploited for preclinical studies on T1D under experimental conditions of synchronized, advanced β cell autoimmunity. Overall, our studies highlight the continuous requirement of Foxp3+ Treg cell activity for the control of genetically pre-installed autoimmune diabetes. AU - Watts, D. AU - Janßen, M. AU - Jaykar, M.* AU - Palmucci, F. AU - Weigelt, M.* AU - Petzold, C.* AU - Hommel, A.* AU - Sparwasser, T.* AU - Bonifacio, E. AU - Kretschmer, K. C1 - 62866 C2 - 51119 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Transient depletion of Foxp3+ regulatory T cells selectively promotes aggressive β cell autoimmunity in genetically susceptible DEREG mice. JO - Front. Immunol. VL - 12 PB - Frontiers Media Sa PY - 2021 SN - 1664-3224 ER - TY - JOUR AB - Asthma is a heterogeneous disease with increasing prevalence worldwide characterized by chronic airway inflammation, increased mucus secretion and bronchial hyperresponsiveness. The phenotypic heterogeneity among asthmatic patients is accompanied by different endotypes, mainly Type 2 or non-Type 2. To investigate the pathomechanism of this complex disease many animal models have been developed, each trying to mimic specific aspects of the human disease. Rodents have classically been employed in animal models of asthma. The present review provides an overview of currently used Type 2 vs. non-Type 2 rodent asthma models, both acute and chronic. It further assesses the methods used to simulate disease development and exacerbations as well as to quantify allergic airway inflammation, including lung physiologic, cellular and molecular immunologic responses. Furthermore, the employment of genetically modified animals, which provide an in-depth understanding of the role of a variety of molecules, signaling pathways and receptors implicated in the development of this disease as well as humanized models of allergic inflammation, which have been recently developed to overcome differences between the rodent and human immune systems, are discussed. Nevertheless, differences between mice and humans should be carefully considered and limits of extrapolation should be wisely taken into account when translating experimental results into clinical use. AU - Alessandrini, F. AU - Musiol, S. AU - Schneider, E. AU - Blanco-Pérez, F.* AU - Albrecht, M.* C1 - 60332 C2 - 49253 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Mimicking antigen-driven asthma in rodent models-how close can we get? JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Allergic reactions to stings of Hymenoptera species may be severe and are potentially fatal deviations of the immunological response observed in healthy individuals. However, venom-specific immunotherapy (VIT) is an immunomodulatory approach able to cure venom allergy in the majority of affected patients. An appropriate therapeutic intervention and the efficacy of VIT not only depend on a conclusive diagnosis, but might also be influenced by the patient-specific manifestation of the disease. As with other diseases, it should be borne in mind that there are different endotypes and phenotypes of venom allergy, each of which require a patient-tailored disease management and treatment scheme. Reviewed here are different endotypes of sting reactions such as IgE-mediated allergy, asymptomatic sensitization or a simultaneous presence of venom allergy and mast cell disorders including particular considerations for diagnosis and therapy. Additionally, phenotypical manifestations of venom allergy, as e.g. differences in age of onset and disease severity, multiple sensitization or patients unsusceptible to therapy, are described. Moreover, biomarkers and diagnostic strategies that might reflect the immunological status of the patient and their value for therapeutic guidance are discussed. Taken together, the increasing knowledge of different disease manifestations in venom hypersensitivity and the growing availability of diagnostic tools open new options for the classification of venom allergy and, hence, for personalized medical approaches and precision medicine in Hymenoptera venom allergy. AU - Blank, S. AU - Grosch, J. AU - Ollert, M.* AU - Bilò, M.B.* C1 - 60503 C2 - 49352 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Precision medicine in hymenoptera venom allergy: Diagnostics, biomarkers, and therapy of different endotypes and phenotypes. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - The same mechanisms that enable host defense against helminths also drive allergic inflammation. This suggests that pathomechanisms of allergic diseases represent evolutionary old responses against helminth parasites and that studying antihelminth immunity may provide insights into pathomechanisms of asthma. However, helminths have developed an intricate array of immunoregulatory mechanisms to modulate type 2 immune mechanisms. This has led to the hypothesis that the lack of helminth infection may contribute to the rise in allergic sensitization in modern societies. Indeed, the anti-inflammatory potential of helminth (worm) parasites and their products in allergy and asthma has been recognized for decades. As helminth infections bring about multiple undesired effects including an increased susceptibility to other infections, intended helminth infection is not a feasible approach to broadly prevent or treat allergic asthma. Thus, the development of new helminth-based biopharmaceutics may represent a safer approach of harnessing type 2-suppressive effects of helminths. However, progress regarding the mechanisms and molecules that are employed by helminths to modulate allergic inflammation has been relatively recent. The scavenging of alarmins and the modulation of lipid mediator pathways and macrophage function by helminth proteins have been identified as important immunoregulatory mechanisms targeting innate immunity in asthma and allergy. In addition, by regulating the activation of dendritic cells and by promoting regulatory T-cell responses, helminth proteins can counterregulate the adaptive T helper 2 cell response that drives allergic inflammation. Despite these insights, important open questions remain to be addressed before helminth molecules can be used for the prevention and treatment of asthma and other allergic diseases. AU - Bohnacker, S. AU - Troisi, F. AU - de los Reyes Jimenez, M. AU - Esser-von Bieren, J. C1 - 60167 C2 - 49284 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - What can parasites tell us about the pathogenesis and treatment of asthma and allergic diseases? JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - In spite of intensive treatment Type 1 diabetes leads to serious complications. Preservation of residual beta cell function makes the disease milder, facilitates treatment, prevents complications and increase survival. So far immune interventions have had limited effect, and some serious adverse events and risks. In an open pilot trial we aimed to improve efficacy of GAD-alum treatment using lymph-node administration in combination with oral vitamin D. Here we report the clinical effect and focus on biomarkers for response to treatment. Patients (n = 12) aged 12 to 24 years with recent onset of Type 1 diabetes received 4 mu g GAD-alum into lymph-node at day 30, 60, and 90, and oral Vitamin D 2000 U/d, days 1 to 120. Beta cell function was estimated by Mixed Meal Tolerance Tests. GADA, GADA subclasses, GAD(65)-induced cytokines and proliferation, and T cells markers were analyzed. The treatment was tolerable with no adverse events. Fasting C-peptide and insulin requirement remained stable at 15 months, while HbA1c was lower than baseline. Stimulated C-peptide showed no change at 6 months but declined after 15 months (81% of baseline). Eleven patients remained in partial remission (IDAAC < 9). Patients (n = 9) with better clinical outcome had reduced proportion of IgG1 and increased IgG2, IgG3, and IgG4, increased IL-10 secretion, and reduction of proliferation and CD8(+) T cells activation. Patients with poorer clinical response had higher baseline levels of GAD(65-)induced cytokines and T-cell activation, and an increased ratio of effector/central memory T cells. Intra-lymphatic GAD treatment combined with Vitamin D might preserve beta cell function and improve clinical course in T1D. Patients with less benefit have a different quality of immune response both before and after treatment. AU - Casas, R.* AU - Dietrich, F.* AU - Barcenilla, H.* AU - Tavira, B.* AU - Wahlberg, J.* AU - Achenbach, P. AU - Ludvigsson, J.* C1 - 60440 C2 - 49316 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Glutamic acid decarboxylase injection into lymph nodes: Beta cell function and immune responses in recent onset type 1 diabetes patients. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - The molecular mechanisms driving specific regulation of neutrophils are not completely understood to date. In order to characterize fundamental granulocyte features on protein level, we analyzed changes in proteome composition as reaction to stress from cell activation processes. For this purpose, we isolated primary granulocytes from equine whole blood through density gradient centrifugation followed by sodium chloride lysis and stimulated cells for 30 min with interleukin-8 (IL8) due to its role as a chemotactic factor for neutrophils. We additionally used phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharide (LPS), which are primarily associated to neutrophil extracellular trap formation and release of reactive oxygen species. From mass spectrometry analysis, we identified a total of 2,032 proteins describing the whole granulocyte proteome, including 245 proteins (12% of identified proteome) newly associated to in vivo expression in primary equine granulocytes (hypothetical proteins). We also found distinct and different changes in protein abundance (ratio >= 2) after short stimulation of cells with various stimuli, pointing to rapid and differentiated reaction pattern. IL8 stimulation resulted in increased protein abundance of 58 proteins (3% of proteome), whereas PMA induced changed protein abundance of 207 (10 % of proteome) and LPS of 46 proteins (2% of proteome). Enrichment analyses clearly showed fundamental differences between stimuli, with primary association of IL8 stimulation to processes in immune response, receptor signaling and signal transduction. Top enrichment for PMA on the other hand pointed to vesicle mediated transport and exocytosis. Stimulation with LPS did not result in any significant enrichment. Although we detected 43% overlap of enrichment categories for IL8 and PMA stimulation, indicating that activation of neutrophils with different stimuli partly induces some similar biological processes and pathways, hierarchical clustering showed clear differences in distribution and biological relevance of clusters between the chosen stimuli. Our studies provide novel information on the granulocyte proteome and offer insights into early, differentiated granulocyte reaction to stimuli, which contribute to a better understanding of molecular mechanisms involved in activation and recruitment of neutrophils, through inflammatory stimuli. AU - Degroote, R.L.* AU - Weigand, M.* AU - Hauck, S.M. AU - Deeg, C.A.* C1 - 58099 C2 - 48201 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - IL8 and PMA trigger the regulation of different biological processes in Granulocyte activation. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Dendritic cells (DC) play a key role in the adaptive immune response due to their ability to present antigens and stimulate naive T cells. Many bacteria and viruses can efficiently target DC, resulting in impairment of their immunostimulatory function or elimination. Hence, the DC compartment requires replenishment following infection to ensure continued operational readiness of the adaptive immune system. Here, we investigated the molecular and cellular mechanisms of inflammation-induced DC generation. We found that infection with viral and bacterial pathogens as well as Toll-like receptor 9 (TLR9) ligation with CpG-oligodeoxynucleotide (CpG-ODN) expanded an erythropoietin (EPO)-dependent TER119(+)CD11a(+)cell population in the spleen that had the capacity to differentiate into TER119(+)CD11c(high)and TER119(-)CD11c(high)cells bothin vitroandin vivo. TER119(+)CD11c(high)cells contributed to the conventional DC pool in the spleen and specifically increased in lymph nodes draining the site of local inflammation. Our results reveal a so far undescribed inflammatory EPO-dependent pathway of DC differentiation and establish a mechanistic link between innate immune recognition of potential immunosuppressive pathogens and the maintenance of the DC pool during and after infection. AU - Einwächter, H.* AU - Heiseke, A.* AU - Schlitzer, A.* AU - Gasteiger, G.* AU - Adler, H. AU - Voehringer, D.* AU - Manz, M.G.* AU - Ruzsics, Z.* AU - Dölken, L.* AU - Koszinowski, U.H.* AU - Sparwasser, T.* AU - Reindl, W.* AU - Jordan, S.* C1 - 59945 C2 - 48947 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - The innate immune response to infection induces erythropoietin-dependent replenishment of the dendritic cell compartment. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Allergic bronchial asthma is a chronic disease of the airways that is characterized by symptoms like respiratory distress, chest tightness, wheezing, productive cough, and acute episodes of broncho-obstruction. This symptom-complex arises on the basis of chronic allergic inflammation of the airway wall. Consequently, the airway epithelium is central to the pathogenesis of this disease, because its multiple abilities directly have an impact on the inflammatory response and thus the formation of the disease. In turn, its structure and functions are markedly impaired by the inflammation. Hence, the airway epithelium represents a sealed, self-cleaning barrier, that prohibits penetration of inhaled allergens, pathogens, and other noxious agents into the body. This barrier is covered with mucus that further contains antimicrobial peptides and antibodies that are either produced or specifically transported by the airway epithelium in order to trap these particles and to remove them from the body by a process called mucociliary clearance. Once this first line of defense of the lung is overcome, airway epithelial cells are the first cells to get in contact with pathogens, to be damaged or infected. Therefore, these cells release a plethora of chemokines and cytokines that not only induce an acute inflammatory reaction but also have an impact on the alignment of the following immune reaction. In case of asthma, all these functions are impaired by the already existing allergic immune response that per se weakens the barrier integrity and self-cleaning abilities of the airway epithelium making it more vulnerable to penetration of allergens as well as of infection by bacteria and viruses. Recent studies indicate that the history of allergy- and pathogen-derived insults can leave some kind of memory in these cells that can be described as imprinting or trained immunity. Thus, the airway epithelium is in the center of processes that lead to formation, progression and acute exacerbation of asthma. AU - Frey, A.* AU - Lunding, L.P.* AU - Ehlers, J.C.* AU - Weckmann, M.* AU - Zissler, U.M. AU - Wegmann, M.* C1 - 59123 C2 - 48575 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - More than just a barrier: The immune functions of the airway epithelium in asthma pathogenesis. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - People with diabetes mellitus have an increased risk for infections, however, there is still a critical gap in precise knowledge about altered immune mechanisms in this disease. Since diabetic INSC94Y transgenic pigs exhibit elevated blood glucose and a stable diabetic phenotype soon after birth, they provide a favorable model to explore functional alterations of immune cells in an early stage of diabetes mellitus in vivo. Hence, we investigated peripheral blood mononuclear cells (PBMC) of these diabetic pigs compared to non-diabetic wild-type littermates. We found a 5-fold decreased proliferative response of T cells in INSC94Y tg pigs to polyclonal T cell mitogen phytohemagglutinin (PHA). Using label-free LC-MS/MS, a total of 3,487 proteins were quantified, and distinct changes in protein abundances in CD4+ T cells of early-stage diabetic pigs were detectable. Additionally, we found significant increases in mitochondrial oxygen consumption rate (OCR) and higher basal glycolytic activity in PBMC of diabetic INSC94Y tg pigs, indicating an altered metabolic immune cell phenotype. Thus, our study provides new insights into molecular mechanisms of dysregulated immune cells triggered by permanent hyperglycemia. AU - Giese, I.M.* AU - Schilloks, M.C.* AU - Degroote, R.L.* AU - Weigand, M.* AU - Renner, S.* AU - Wolf, E.* AU - Hauck, S.M. AU - Deeg, C.A.* C1 - 61333 C2 - 50130 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Chronic hyperglycaemia drives functional impairment of lymphocytes in diabetic INSC94Y transgenic pigs. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Currently three bona fide dendritic cell (DC) types are distinguished in human blood. Herein we focus on type 2 DCs (DC2s) and compare the three defining markers CD1c, CD172, and CD301. When using CD1c to define DC2s, a CD14(+)and a CD14(-)subset can be detected. The CD14(+)subset shares features with monocytes, and this includes substantially higher expression levels for CD64, CD115, CD163, and S100A8/9. We review the current knowledge of these CD1c(+)CD14(+)cells as compared to the CD1c(+)CD14(-)cells with respect to phenotype, function, transcriptomics, and ontogeny. Here, we discuss informative mutations, which suggest that two populations have different developmental requirements. In addition, we cover subsets of CD11c(+)CD8(-)DC2s in the mouse, where CLEC12A(+)ESAM(low)cells, as compared to the CLEC12A(-)ESAM(high)subset, also express higher levels of monocyte-associated markers CD14, CD3, and CD115. Finally, we summarize, for both man and mouse, the data on lower antigen presentation and higher cytokine production in the monocyte-marker expressing DC2 subset, which demonstrate that the DC2 subsets are also functionally distinct. AU - Heger, L.* AU - Hofer, T.P. AU - Bigley, V.* AU - de Vries, I.J.M.* AU - Dalod, M.* AU - Dudziak, D.* AU - Ziegler-Heitbrock, L.* C1 - 60338 C2 - 49208 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Subsets of CD1c+DCs: Dendritic cell versus monocyte lineage. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Although first described decades ago, the relevance of carbohydrate specific antibodies as mediators of type I allergy had not been recognized until recently. Previously, allergen specific IgE antibodies binding to carbohydrate epitopes were considered to demonstrate a clinically irrelevant cross-reactivity. However, this changed following the discovery of type I allergies specifically mediated by oligosaccharide structures. Especially the emerging understanding of red meat allergy characterized by IgE directed to the oligosaccharide alpha-gal showed that carbohydrate-mediated reactions can result in life threatening systemic anaphylaxis which in contrast to former assumptions proves a high clinical relevance of some carbohydrate allergens. Within the scope of this review article, we illustrate the historical development of carbohydrate-allergen-research, reaching from only diagnostically relevant crossreactive-carbohydrate-determinants to clinically important antigens mediating type I allergy. Focusing on clinical and immunological features of the alpha-gal syndrome, we highlight the discovery of oligosaccharides as potentially highly immunogenic antigens and mediators of type I allergy, report what is known about the route of sensitization and the immunological mechanisms involved in sensitization and elicitation phase of allergic responses as well as currently available diagnostic and therapeutic tools. Finally, we briefly report on carbohydrates being involved in type I allergies different from alpha-gal. AU - Hils, M.* AU - Wölbing, F.* AU - Hilger, C.* AU - Fischer, J.* AU - Hoffard, N.* AU - Biedermann, T. C1 - 60432 C2 - 49329 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - The history of carbohydrates in type I allergy. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Food allergy is an atopic disease that is caused by the immune system targeting harmless food antigens that can result in life-threatening anaphylaxis. As humans and microbes have co-evolved, inevitably commensal microbes have a tremendous impact on our health. As such, the gut with its enormous microbial richness reflects a highly tolerogenic environment at steady state, in which immune cells are educated to react in a well-calibrated manner to food and microbial antigens. Recent evidence indicates that the susceptibility to food allergy is critically linked to microbial dysbiosis and can be transmitted by microbial transfer from humans to mice. Experimental work and epidemiological studies further point toward a critical time window in early childhood during which the immune system is imprinted by microbial colonization. Particularly, Foxp3-expressing regulatory T cells turn out to be key players, acting as rheostats for controlling the magnitude of food allergic reactions. An increasing number of bacterial metabolites has recently been shown to regulate directly or indirectly the differentiation of peripherally induced Tregs, most of which co-express the RAR-related orphan receptor gamma t (ROR gamma t). Genetic ablation provided additional direct evidence for the importance of ROR gamma t+ Tregs in food allergy. Future strategies for the stratification of food allergic patients with the aim to manipulate the intestinal microbiota by means of fecal transplantation efforts, pre- or probiotic regimens or for boosting oral immunotherapy may improve diagnosis and therapy. In this review some of the key underlying mechanisms are summarized and future directions for potential microbial therapy are explored. AU - Kreft, L. AU - Hoffmann, C. AU - Ohnmacht, C. C1 - 59999 C2 - 49160 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Therapeutic potential of the intestinal microbiota for immunomodulation of food allergies. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - BackgroundSevere Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) induced Coronavirus Disease 2019 (COVID-19) has posed a global threat to public health. The immune system is crucial in defending and eliminating the virus and infected cells. However, immune dysregulation may result in the rapid progression of COVID-19. Here, we evaluated the subsets, phenotypic and functional characteristics of natural killer (NK) and T cells in patients with COVID-19 and their associations with disease severity.MethodsDemographic and clinical data of COVID-19 patients enrolled in Wuhan Union Hospital from February 25 to February 27, 2020, were collected and analyzed. The phenotypic and functional characteristics of NK cells and T cells subsets in circulating blood and serum levels of cytokines were analyzed via flow cytometry. Then the LASSO logistic regression model was employed to predict risk factors for the severity of COVID-19.ResultsThe counts and percentages of NK cells, CD4(+) T cells, CD8(+) T cells and NKT cells were significantly reduced in patients with severe symptoms. The cytotoxic CD3(-)CD56(dim)CD16(+) cell population significantly decreased, while the CD3(-)CD56(dim)CD16(-) part significantly increased in severe COVID-19 patients. More importantly, elevated expression of regulatory molecules, such as CD244 and programmed death-1 (PD-1), on NK cells and T cells, as well as decreased serum cytotoxic effector molecules including perforin and granzyme A, were detected in patients with COVID-19. The serum IL-6, IL-10, and TNF-alpha were significantly increased in severe patients. Moreover, the CD3(-)CD56(dim)CD16(-) cells were screened out as an influential factor in severe cases by LASSO logistic regression.ConclusionsThe functional exhaustion and other subset alteration of NK and T cells may contribute to the progression and improve the prognosis of COVID-19. Surveillance of lymphocyte subsets may in the future enable early screening for signs of critical illness and understanding the pathogenesis of this disease. AU - Li, M.* AU - Guo, W.* AU - Dong, Y.* AU - Wang, X.* AU - Dai, D.* AU - Liu, X.* AU - Wu, Y.* AU - Zhang, W.* AU - Zhou, H.* AU - Zhang, Z.* AU - Lin, L.* AU - Kang, Z.* AU - Yu, T.* AU - Tian, C.* AU - Qin, R.* AU - Gui, Y.* AU - Jiang, F.* AU - Fan, H.* AU - Heissmeyer, V. AU - Sarapultsev, A.* AU - Wang, L.* AU - Luo, S.* AU - Hu, D.* C1 - 60448 C2 - 49461 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Elevated exhaustion levels of NK and CD8+ T cells as indicators for progression and prognosis of COVID-19 disease. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - The pseudokinase TRIB1 controls cell function in a range of contexts, by regulating MAP kinase activation and mediating protein degradation via the COP1 ubiquitin ligase. TRIB1 regulates polarization of macrophages and dysregulated Trib1 expression in murine models has been shown to alter atherosclerosis burden and adipose homeostasis. Recently, TRIB1 has also been implicated in the pathogenesis of prostate cancer, where it is often overexpressed, even in the absence of genetic amplification. Well described TRIB1 effectors include MAP kinases and C/EBP transcription factors, both in immune cells and in carcinogenesis. However, the mechanisms that regulate TRIB1 itself remain elusive. Here, we show that the long and conserved 3’untranslated region (3’UTR) of TRIB1 is targeted by miRNAs in macrophage and prostate cancer models. By using a systematic in silico analysis, we identified multiple “high confidence” miRNAs potentially binding to the 3’UTR of TRIB1 and report that miR-101-3p and miR-132-3p are direct regulators of TRIB1 expression and function. Binding of miR-101-3p and miR-132-3p to the 3’UTR of TRIB1 mRNA leads to an increased transcription and secretion of interleukin-8. Our data demonstrate that modulation of TRIB1 by miRNAs alters the inflammatory profile of both human macrophages and prostate cancer cells. AU - Niespolo, C.* AU - Johnston, J.M.* AU - Deshmukh, S.R.* AU - Satam, S. AU - Shologu, Z.* AU - Villacanas, O.* AU - Sudbery, I.M.* AU - Wilson, H.L.* AU - Kiss-Toth, E.* C1 - 60821 C2 - 49655 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Tribbles-1 expression and its function to control inflammatory cytokines, including interleukin-8 levels are regulated by miRNAs in macrophages and prostate cancer cells. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - Double negative (DN) (CD19+CD20lowCD27-IgD-) B cells are expanded in patients with autoimmune and infectious diseases; however their role in the humoral immune response remains unclear. Using systematic flow cytometric analyses of peripheral blood B cell subsets, we observed an inflated DN B cell population in patients with variety of active inflammatory conditions: myasthenia gravis, Guillain-Barré syndrome, neuromyelitis optica spectrum disorder, meningitis/encephalitis, and rheumatic disorders. Furthermore, we were able to induce DN B cells in healthy subjects following vaccination against influenza and tick borne encephalitis virus. Transcriptome analysis revealed a gene expression profile in DN B cells that clustered with naïve B cells, memory B cells, and plasmablasts. Immunoglobulin VH transcriptome sequencing and analysis of recombinant antibodies revealed clonal expansion of DN B cells that were targeted against the vaccine antigen. Our study suggests that DN B cells are expanded in multiple inflammatory neurologic diseases and represent an inducible B cell population that responds to antigenic stimulation, possibly through an extra-follicular maturation pathway. AU - Ruschil, C.* AU - Gabernet, G.* AU - Lepennetier, G.* AU - Heumos, S.* AU - Kaminski, M.* AU - Hracsko, Z.* AU - Irmler, M. AU - Beckers, J. AU - Ziemann, U.* AU - Nahnsen, S.* AU - Owens, G.P.* AU - Bennett, J.L.* AU - Hemmer, B.* AU - Kowarik, M.C.* C1 - 60919 C2 - 49631 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Specific induction of double negative B cells during protective and pathogenic immune responses. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - The immune response to antigens is a key aspect of immunology, as it provides opportunities for therapeutic intervention. However, the induction of immunological tolerance is an evolving area that is still not sufficiently understood. Allergen immunotherapy (AIT) is a disease-modulating therapy available for immunoglobulin E (IgE)-mediated airway diseases such as allergic rhinitis or allergic asthma. This disease-modifying effect is not only antigen driven but also antigen specific. The specificity and also the long-lasting, often life-long symptom reduction make the therapy attractive for patients. Additionally, the chance to prevent the onset of asthma by treating allergic rhinitis with AIT is important. The mechanism and, in consequence, therapy guiding biomarker are still in its infancy. Recent studies demonstrated that the interaction of T, B, dendritic, and epithelial cells and macrophages are individually contributing to clinical tolerance and therefore underline the need for a system to monitor the progress and success of AIT. As clinical improvement is often accompanied by decreases in numbers of effector cells in the tissue, analyses of cellular responses and cytokine pattern provide a good insight into the mechanisms of AIT. The suppression of type-2 immunity is accompanied by decreased levels of type-2 mediators such as epithelial CCL-26 and interleukin (IL)-4, IL-13 produced by T cells that are constituting the immune memory and are increasingly controlled by regulatory T and B cells following AIT. Immune tolerance is also associated with increased production of type-1 mediators like interferon-gamma, tissue-homeostating factors like indoleamine 2,3-dioxygenase (IDO) expressed by macrophages and dendritic cells. Although these individual genes were convincingly demonstrated to play a role immune tolerance, they do not predict therapy outcomes of AIT on an individual level. Therefore, combinations or ratios of gene expression levels are a promising way to achieve predictive value and definition of helpful biomarker. AU - Zissler, U.M. AU - Schmidt-Weber, C.B. C1 - 60059 C2 - 49200 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Predicting success of allergen-specific immunotherapy. JO - Front. Immunol. VL - 11 PB - Frontiers Media Sa PY - 2020 SN - 1664-3224 ER - TY - JOUR AB - For many decades, glucocorticoids have been widely used as the gold standard treatment for inflammatory conditions. Unfortunately, their clinical use is limited by severe adverse effects such as insulin resistance, cardiometabolic diseases, muscle and skin atrophies, osteoporosis, and depression. Glucocorticoids exert their effects by binding to the Glucocorticoid Receptor (GR), a ligand-activated transcription factor which both positively, and negatively regulates gene expression. Extensive research during the past several years has uncovered novel mechanisms by which the GR activates and represses its target genes. Genome-wide studies and mouse models have provided valuable insight into the molecular mechanisms of inflammatory gene regulation by GR. This review focusses on newly identified target genes and GR co-regulators that are important for its anti-inflammatory effects in innate immune cells, as well as mutations within the GR itself that shed light on its transcriptional activity. This research progress will hopefully serve as the basis for the development of safer immune suppressants with reduced side effect profiles. AU - Escoter Torres, L. AU - Caratti, G.* AU - Mechtidou, A. AU - Tuckermann, J.* AU - Uhlenhaut, N.H. AU - Vettorazzi, S.* C1 - 56785 C2 - 47293 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Fighting the fire: Mechanisms of inflammatory gene regulation by the glucocorticoid receptor. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Autoantibodies to myelin oligodendrocytes glycoprotein (MOG) are found in a fraction of patients with inflammatory demyelination and are detected with MOG-transfected cells. While the prototype anti-MOG mAb 8-18C5 and polyclonal anti-MOG responses from different mouse strains largely recognize the FG loop of MOG, the human anti-MOG response is more heterogeneous and human MOG-Abs recognizing different epitopes were found to be pathogenic. The aim of this study was to get further insight into details of antigen-recognition by human MOG-Abs focusing on the impact of glycosylation. MOG has one known N-glycosylation site at N31 located in the BC loop linking two beta-sheets. We compared the reactivity to wild type MOG with that toward two different mutants in which the neutral asparagine of N31 was mutated to negatively charged aspartate or to the neutral alanine. We found that around 60% of all patients (16/27) showed an altered reactivity to one or both of the mutations. We noted seven different patterns of recognition of the two glycosylation-deficient mutants by different patients. The introduced negative charge at N31 enhanced recognition in some, but reduced recognition in other patients. In 7/27 patients the neutral glycosylation-deficient mutant was recognized stronger. The folding of the extracellular domain of MOG with the formation of beta-sheets did not depend on its glycosylation as seen by circular dichroism. We determined the glycan structure of MOG produced in HEK cells by mass spectrometry. The most abundant glycoforms of MOG expressed in HEK cells are diantennary, contain a core fucose, an antennary fucose, and are decorated with α2,6 linked Neu5Ac, while details of the glycoforms of MOG in myelin remain to be identified. Together, we (1) increase the knowledge about heterogeneity of human autoantibodies to MOG, (2) show that the BC loop affects recognition in about 60% of the patients, (3) report that all patients recognized the unglycosylated protein backbone, while (4) in about 20% of the patients the attached sugar reduces autoantibody binding presumably via steric hindrance. Thus, a neutral glycosylation-deficient mutant of MOG might enhance the sensitivity to identify MOG-Abs. AU - Fernandez, I.M.* AU - Macrini, C.* AU - Krumbholz, M.* AU - Hensbergen, P.J.* AU - Ederveen, A.L.H.* AU - Winklmeier, S.* AU - Vural, A.* AU - Kurne, A.* AU - Jenne, D. AU - Kamp, F.* AU - Gerdes, L.A.* AU - Hohlfeld, R.* AU - Wuhrer, M.* AU - Kuempfel, T.* AU - Meinl, E.* C1 - 56401 C2 - 47061 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - The glycosylation site of myelin oligodendrocyte glycoprotein affects autoantibody recognition in a large proportion of patients. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - CD8(+) T cells are important effectors of adaptive immunity against pathogens, tumors, and self antigens. Here, we asked how human cognate antigen-responsive CD8(+) T cells and their receptors could be identified in unselected single-cell gene expression data. Single-cell RNA sequencing and qPCR of dye-labeled antigen-specific cells identified large gene sets that were congruently up- or downregulated in virus-responsive CD8(+) T cells under different antigen presentation conditions. Combined expression of TNFRSF9, XCL1, XCL2, and CRTAM was the most distinct marker of virus-responsive cells on a single-cell level. Using transcriptomic data, we developed a machine learning-based classifier that provides sensitive and specific detection of virus-responsive CD8(+) T cells from unselected populations. Gene response profiles of CD8(+) T cells specific for the autoantigen islet-specific glucose-6-phosphatase catalytic subunit-related protein differed markedly from virus-specific cells. These findings provide single-cell gene expression parameters for comprehensive identification of rare antigen-responsive cells and T cell receptors. AU - Fuchs, Y.F.* AU - Sharma, V.* AU - Eugster, A.* AU - Kraus, G.* AU - Morgenstern, R.* AU - Dahl, A.* AU - Reinhardt, S.* AU - Petzold, A.* AU - Lindner, A.* AU - Löbel, D.* AU - Bonifacio, E. C1 - 57465 C2 - 47791 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Gene expression-based identification of antigen-responsive CD8+ T cells on a single-cell level. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Copyright © 2019 Hofer, van de Loosdrecht, Stahl-Hennig, Cassatella and Ziegler-Heitbrock. Monocytes are subdivided into three subsets, which have different phenotypic and functional characteristics and different roles in inflammation and malignancy. When in man CD14 and CD16 monoclonal antibodies are used to define these subsets, then the distinction of non-classical CD14low and intermediate CD14high monocytes requires setting a gate in what is a gradually changing level of CD14 expression. In the search for an additional marker to better dissect the two subsets we have explored the marker 6-sulfo LacNAc (slan). Slan is a carbohydrate residue originally described to be expressed on the cell surface of a type of dendritic cell in human blood. We elaborate herein that the features of slan+ cells are congruent with the features of CD16+ non-classical monocytes and that slan is a candidate marker for definition of non-classical monocytes. The use of this marker may help in studying the role of non-classical monocytes in health and in diagnosis and monitoring of disease. AU - Hofer, T.P. AU - van de Loosdrecht, A.* AU - Stahl-Hennig, C.* AU - Cassatella, M.A.* AU - Ziegler-Heitbrock, L.* C1 - 56917 C2 - 47280 TI - 6-sulfo LacNAc (Slan) as a marker for non-classical monocytes. JO - Front. Immunol. VL - 10 PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Influenza vaccination is a common approach to prevent seasonal and pandemic influenza. Pre-existing antibodies against close viral strains might impair antibody formation against previously unseen strains-a process called original antigenic sin. The role of this pre-existing cellular immunity in this process is, despite some hints from animal models, not clear. Here, we analyzed cellular and humoral immunity in healthy individuals before and after vaccination with seasonal influenza vaccine. Based on influenza-specific hemagglutination inhibiting (HI) titers, vaccinees were grouped into HI-negative and -positive cohorts followed by in-depth cytometric and TCR repertoire analysis. Both serological groups revealed cross-reactive T-cell memory to the vaccine strains at baseline that gave rise to the majority of vaccine-specific T-cells post vaccination. On the contrary, very limited number of vaccine-specific T-cell clones was recruited from the naive pool. Furthermore, baseline quantity of vaccine-specific central memory helper T-cells and clonotype richness of this population directly correlated with the vaccination efficacy. Our findings suggest that the deliberate recruitment of pre-existing cross-reactive cellular memory might help to improve vaccination outcome. AU - Nienen, M.* AU - Stervbo, U.* AU - Moelder, F.* AU - Kaliszczyk, S.* AU - Kuchenbecker, L.* AU - Gayova, L.* AU - Schweiger, B.* AU - Juerchott, K.* AU - Hecht, J.* AU - Neumann, A.U. AU - Rahmann, S.* AU - Westhoff, T.H.* AU - Reinke, P.* AU - Thiel, A.* AU - Babel, N.* C1 - 55879 C2 - 46642 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - The role of pre-existing cross-reactive central memory CD4 T-cells in vaccination with previously unseen influenza strains. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Mutants of a catalytically inactive variant of Proteinase 3 (PR3)-iPR3-Val(103) possessing a Ser195Ala mutation relative to wild-type PR3-Val(103)-offer insights into how autoantigen PR3 interacts with antineutrophil cytoplasmic antibodies (ANCAs) in granulomatosis with polyangiitis (GPA) and whether such interactions can be interrupted. Here we report that iHm5-Val(103), a triple mutant of iPR3-Val(103), bound a monoclonal antibody (moANCA518) from a GPA patient on an epitope remote from the mutation sites, whereas the corresponding epitope of iPR3-Val(103) was latent to moANCA518. Simulated B-factor analysis revealed that the binding of moANCA518 to iHm5-Val(103) was due to increased main-chain flexibility of the latent epitope caused by remote mutations, suggesting rigidification of epitopes with therapeutics to alter pathogenic PR3 center dot ANCA interactions as new GPA treatments. AU - Pang, Y.P.* AU - Moura, M.C.* AU - Thompson, G.E.* AU - Nelson, D.R.* AU - Hummel, A.M.* AU - Jenne, D. AU - Emerling, D.* AU - Volkmuth, W.* AU - Robinson, W.H.* AU - Specks, U.* C1 - 57313 C2 - 47713 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Remote activation of a latent epitope in an autoantigen decoded with simulated B-factors. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Recent advances in cytometry have radically altered the fate of single-cell proteomics by allowing a more accurate understanding of complex biological systems. Mass cytometry (CyTOF) provides simultaneous single-cell measurements that are crucial to understand cellular heterogeneity and identify novel cellular subsets. High-dimensional CyTOF data were traditionally analyzed by gating on bivariate dot plots, which are not only laborious given the quadratic increase of complexity with dimension but are also biased through manual gating. This review aims to discuss the impact of new analysis techniques for in-depths insights into the dynamics of immune regulation obtained from static snapshot data and to provide tools to immunologists to address the high dimensionality of their single-cell data. AU - Patil, S.* AU - Heuser, C.* AU - de Almeida, G.P.* AU - Theis, F.J. AU - Zielinski, C.E.* C1 - 56587 C2 - 47109 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Meeting the challenges of high-dimensional single-cell data analysis in immunology. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Gammaherpesviruses (gamma HV) are important pathogens causing persistent infections which lead to several malignancies in immunocompromised patients. Murine gamma HV 68 (MHV-68), a homolog to human EBV and KSHV, has been employed as a classical pathogen to investigate the molecular pathogenicity of gamma HV infections. gamma HV express distinct antigens during lytic or latent infection and antigen-specific T cells have a significant role in controlling the acute and latent viral infection, although the quality of anti-viral T cell responses required for protective immunity is not well-understood. We have generated recombinant modified vaccinia virus Ankara (recMVA) vaccines via MVA-BAC homologous recombination technology expressing MHV-68 ORF6 and ORF61 antigens encoding both MHC class I and II-restricted epitopes. After vaccination, we examined T cell responses before and after MHV-68 infection to determine their involvement in latent virus control. We show recognition of recMVA- and MHV-68-infected APC by ORF6 and ORF61 epitope-specific T cell lines in vitro. The recMVA vaccines efficiently induced MHV-68-specific CD8+ and CD4+ T cell responses after a single immunization and more pronounced after homologous prime/boost vaccination in mice. Moreover, we exhibit protective capacity of prophylactic recMVA vaccination during early latency at day 17 after intranasal challenge with MHV-68, but failed to protect from latency at day 45. Further T cell analysis indicated that T cell exhaustion was not responsible for the lack of protection by recMVA vaccination in long-termlatency at day 45. The data support further efforts aiming at improved vaccine development against gamma HV infections with special focus on targeting protective CD4+ T cell responses. AU - Samreen, B.* AU - Tao, S.* AU - Tischer, K.* AU - Adler, H. AU - Drexler, I.* C1 - 57792 C2 - 47920 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - ORF6 and ORF61 expressing MVA vaccines impair early but not late latency in murine gammaherpesvirus MHV-68 infection. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - The IL-7/IL-7R pathway is essential for lymphocyte development and disturbances in the pathway can lead to immune deficiency or T cell mediated destruction. Here, the effect of transient hyperexpression of IL-7 was investigated on immune regulation and allograft rejection under immunosuppression. An experimental in vivo immunosuppressive mouse model of IL-7 hyperexpression was developed using transgenic mice (C57BL/6 background) carrying a tetracycline inducible IL-7 expression cassette, which allowed the temporally controlled induction of IL-7 hyperexpression by Dexamethasone and Doxycycline treatment. Upon induction of IL-7, the B220+ c-kit+ Pro/Pre-B I compartment in the bone marrow increased as compared to control mice in a serum IL-7 concentration-correlated manner. IL-7 hyperexpression also preferentially increased the population size of memory CD8+ T cells in secondary lymphoid organs, and reduced the proportion of CD4+Foxp3+ T regulatory cells. Of relevance to disease, conventional CD4+ T cells from an IL-7-rich milieu escaped T regulatory cell-mediated suppression in vitro and in a model of autoimmune diabetes in vivo. These findings were validated using an IL-7/anti-IL7 complex treatment mouse model to create an IL-7 rich environment. To study the effect of IL-7 on islet graft survival in a mismatched allograft model, BALB/c mice were rendered diabetic by streptozotocin und transplanted with IL-7-inducible or control islets from C57BL/6 mice. As expected, Dexamethasone and Doxycycline treatment prolonged graft median survival as compared to the untreated control group in this transplantation mouse model. However, upon induction of local IL-7 hyperexpression in the transplanted islets, graft survival time was decreased and this was accompanied by an increased CD4+ and CD8+ T cell infiltration in the islets. Altogether, the findings show that transient elevations of IL-7 can impair immune regulation and lead to graft loss also under immune suppression. AU - Schreiber, M.* AU - Weigelt, M. AU - Karasinsky, A.* AU - Anastassiadis, K.* AU - Schallenberg, S.* AU - Petzold, C.* AU - Bonifacio, E. AU - Kretschmer, K. AU - Hommel, A. C1 - 55930 C2 - 46720 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Inducible IL-7 hyperexpression influences lymphocyte homeostasis and function and increases allograft rejection. JO - Front. Immunol. VL - 10 PB - Frontiers Media Sa PY - 2019 SN - 1664-3224 ER - TY - JOUR AB - Aicardi-Goutières syndrome (AGS) is a rare early onset childhood encephalopathy caused by persistent neuroinflammation of autoimmune origin. AGS is a genetic disorder and >50% of affected individuals bear hypomorphic mutations in ribonuclease H2 (RNase H2). All available RNase H2 mouse models so far fail to mimic the prominent CNS involvement seen in AGS. To establish a mouse model recapitulating the human disease, we deleted RNase H2 specifically in the brain, the most severely affected organ in AGS. Although RNase H2δGFAPmice lacked the nuclease in astrocytes and a majority of neurons, no disease signs were apparent in these animals. We additionally confirmed these results in a second, neuron-specific RNase H2 knockout mouse line. However, when astrocytes were isolated from brains of RNase H2δGFAPmice and cultured under mitogenic conditions, they showed signs of DNA damage and premature senescence. Enhanced expression of interferon-stimulated genes (ISGs) represents the most reliable AGS biomarker. Importantly, primary RNase H2δGFAPastrocytes displayed significantly increased ISG transcript levels, which we failed to detect in in vivo in brains of RNase H2δGFAPmice. Isolated astrocytes primed by DNA damage, including RNase H2-deficiency, exhibited a heightened innate immune response when exposed to bacterial or viral antigens. Taken together, we established a valid cellular AGS model that utilizes the very cell type responsible for disease pathology, the astrocyte, and phenocopies major molecular defects observed in AGS patient cells. AU - Bartsch, K.* AU - Damme, M.* AU - Regen, T.* AU - Becker, L. AU - Garrett, L. AU - Hölter, S.M. AU - Knittler, K.* AU - Borowski, C.* AU - Waisman, A.* AU - Glatzel, M.* AU - Fuchs, H. AU - Gailus-Durner, V. AU - Hrabě de Angelis, M. AU - Rabe, B.* C1 - 53382 C2 - 44542 CY - Lausanne TI - RNase H2 loss in murine astrocytes results in cellular defects reminiscent of nucleic acid-mediated autoinflammation. JO - Front. Immunol. VL - 9 IS - MAR PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - T follicular helper (Tfh) cells are critically involved in the establishment of potent antibody responses against infectious pathogens, such as viruses and bacteria, but their dysregulation may also result in aberrant antibody responses that frequently coincide with autoimmune diseases or allergies. The fate and identity of Tfh cells is tightly controlled by gene regulation on the transcriptional and posttranscriptional level. Here, we provide deeper insights into the posttranscriptional mechanisms that regulate Tfh cell differentiation, function, and plasticity through the actions of RNA-binding proteins (RBPs) and small endogenously expressed regulatory RNAs called microRNAs (miRNAs). The Roquin family of RBPs has been shown to dampen spontaneous activation and differentiation of naive CD4(+) T cells into Tfh cells, since CD4(+) T cells with Roquin mutations accumulate as Tfh cells and provide inappropriate B cell help in the production of autoantibodies. Moreover, Regnase-1, an endoribonuclease that regulates a set of targets, which strongly overlaps with that of Roquin, is crucial for the prevention of autoantibody production. Interestingly, both Roquin and Regnase-1 proteins are cleaved and inactivated after TCR stimulation by the paracaspase MALT1. miRNAs are expressed in naive CD4(+) T cells and help preventing spontaneous differentiation into effector cells. While most miRNAs are downregulated upon T cell activation, several miRNAs have been shown to regulate the fate of these cells by either promoting (e.g., miR-17-92 and miR-155) or inhibiting (e.g., miR-146a) Tfh cell differentiation. Together, these different aspects highlight a complex and dynamic regulatory network of posttranscriptional gene regulation in Tfh cells that may also be active in other T helper cell populations, including Th1, Th2, Th17, and Treg. AU - Baumjohann, D.* AU - Heissmeyer, V. C1 - 54103 C2 - 45307 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Posttranscriptional gene regulation of T follicular helper cells by RNA-binding proteins and microRNAs. JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Under physiological conditions, CD4(+) regulatory T (Treg) cells expressing the transcription factor Foxp3 are generated in the thymus [thymus-derived Foxp3(+) Treg (tTregs) cells] and extrathymically at peripheral sites [peripherally induced Foxp3(+) Treg (pTreg) cell], and both developmental subsets play non-redundant roles in maintaining self-tolerance throughout life. In addition, a variety of experimental in vitro and in vivo modalities can extrathymically elicit a Foxp3(+) Treg cell phenotype in peripheral CD4(+) Foxp3(-) T cells, which has attracted much interest as an approach toward cell-based therapy in clinical settings of undesired immune responses. A particularly notable example is the in vitro induction of Foxp3 expression and Treg cell activity (iTreg cells) in initially naive CD4(+)Foxp3(-) T cells through T cell receptor (TCR) and IL-2R ligation, in the presence of exogenous TGF-beta. Clinical application of Foxp3(+) iTreg cells has been hampered by the fact that TGF-beta-driven Foxp3 induction is not sufficient to fully recapitulate the epigenetic and transcriptional signature of in vivo induced Foxp3(+) tTreg and pTreg cells, which includes the failure to imprint iTreg cells with stable Foxp3 expression. This hurdle can be potentially overcome by pharmacological interference with DNA methyltransferase activity and CpG methylation [e.g., by the cytosine nucleoside analog 5-aza-2'-deoxycytidine (5-aza-dC)] to stabilize TGF-beta-induced Foxp3 expression and to promote a Foxp3(+) iTreg cell phenotype even in the absence of added TGF-beta. However, the molecular mechanisms of 5-aza-dC-mediated Foxp3(+) iTreg cell generation have remained incompletely understood. Here, we show that in the absence of exogenously added TGF-beta and IL-2, efficient 5-aza-dC-mediated Foxp3+ iTreg cell generation from TCR-stimulated CD4(+) Foxp3(-) T cells is critically dependent on TGF-beta R and IL-2R signaling and that this process is driven by TGF-beta and IL-2, which could either be FCS derived or produced by T cells on TCR stimulation. Overall, these findings contribute to our understanding of the molecular mechanisms underlying the process of Foxp3 induction and may provide a rational basis for generating phenotypically and functionally stable iTreg cells. AU - Freudenberg, K.* AU - Lindner, N.* AU - Dohnke, S.* AU - Garbe, A.I.* AU - Schallenberg, S.* AU - Kretschmer, K. C1 - 52952 C2 - 44381 CY - Lausanne TI - Critical role of  TGF-β and IL-2 receptor signalling in Foxp3 inducion by an inhibitor of DNA methylation. JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Cellular therapies with CD4+ T regulatory cells (Tregs) hold promise of efficacious treatment for the variety of autoimmune and allergic diseases as well as posttransplant complications. Nevertheless, current manufacturing of Tregs as a cellular medicinal product varies between different laboratories, which in turn hampers precise comparisons of the results between the studies performed. While the number of clinical trials testing Tregs is already substantial, it seems to be crucial to provide some standardized characteristics of Treg products in order to minimize the problem. We have previously developed reporting guidelines called minimum information about tolerogenic antigen-presenting cells, which allows the comparison between different preparations of tolerance-inducing antigen-presenting cells. Having this experience, here we describe another minimum information about Tregs (MITREG). It is important to note that MITREG does not dictate how investigators should generate or characterize Tregs, but it does require investigators to report their Treg data in a consistent and transparent manner. We hope this will, therefore, be a useful tool facilitating standardized reporting on the manufacturing of Tregs, either for research purposes or for clinical application. This way MITREG might also be an important step toward more standardized and reproducible testing of the Tregs preparations in clinical applications. AU - Fuchs, A.* AU - Gliwinski, M.* AU - Grageda, N.* AU - Spiering, R.* AU - Abbas, A.K.* AU - Appel, S.* AU - Bacchetta, R.* AU - Battaglia, M.* AU - Berglund, D.* AU - Blazar, B.* AU - Bluestone, J.A.* AU - Bornhaeuser, M.* AU - ten Brinke, A.* AU - Brusko, T.M.* AU - Cools, N.* AU - Cuturi, M.C.* AU - Geissler, E.* AU - Giannoukakis, N.* AU - Golab, K.* AU - Hafler, D.A.* AU - van Ham, S.M.* AU - Hester, J.* AU - Hippen, K.* AU - Di Ianni, M.* AU - Ilic, N.* AU - Isaacs, J.* AU - Issa, F.* AU - Iwaszkiewicz-Grzes, D.* AU - Jaeckel, E.* AU - Joosten, I.* AU - Klatzmann, D.* AU - Koenen, H.* AU - Van Kooten, C.* AU - Korsgren, O.* AU - Kretschmer, K. AU - Levings, M.* AU - Marek-Trzonkowska, N.M.* AU - Martínez-Llordella, M.* AU - Miljkovic, D.* AU - Mills, K.H.G.* AU - Miranda, J.P.* AU - Piccirillo, C.A.* AU - Putnam, A.L.* AU - Ritter, T.* AU - Roncarolo, M.G.* AU - Sakaguchi, S.* AU - Sanchez-Ramon, S.* AU - Sawitzki, B.* AU - Sofronic-Milosavljevic, L.* AU - Sykes, M.* AU - Tang, Q.* AU - Vives-Pi, M.* AU - Waldmann, H.* AU - Witkowski, P.* AU - Wood, K.J.* AU - Gregori, S.* AU - Hilkens, C.M.U.* AU - Lombardi, G.* AU - Lord, P.* AU - Martinez-Caceres, E.M.* AU - Trzonkowski, P.* C1 - 52847 C2 - 44310 CY - Lausanne TI - Minimum information about T regulatory cells: A step toward reproducibility and standardization. JO - Front. Immunol. VL - 8 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Since the B-cell lymphoma/leukemia 10 (BCL10) protein was first described in 1999, numerous studies have elucidated its key functions in channeling adaptive and innate immune signaling downstream of CARMA/caspase-recruitment domain (CARD) scaffold proteins. While T and B cell antigen receptor (TCR/BCR) signaling induces the recruitment of BCL10 bound to mucosa-associated lymphoid tissue (MALT) 1 to the lymphocyte-specific CARMA1/CARD11-BCL10-MALT1 (CBM-1) signalosome, alternative CBM complexes utilize different CARMA/CARD scaffolds in distinct innate or inflammatory pathways. BCL10 constitutes the smallest subunit in all CBM signalosomes, containing a 233 amino acid coding for N-terminal CARD as well as a C-terminal Ser/Thr-rich region. BCL10 forms filaments, thereby aggregating into higher-order clusters that mediate and amplify stimulation-induced signals, ultimately leading to MALT1 protease activation and canonical NF-kappa B and JNK signaling. BCL10 additionally undergoes extensive post-translational regulation involving phosphorylation, ubiquitination, MALT1-catalyzed cleavage, and degradation. Through these feedback and feed-forward events, BCL10 integrates positive and negative regulatory processes that govern the function as well as the dynamic assembly, disassembly, and destruction of CBM complexes. Thus, BCL10 is a critical regulator for activation as well as termination of immune cell signaling, revealing that its role extends far beyond that of a mere linking factor in CBM complexes. AU - Gehring, T. AU - Seeholzer, T. AU - Krappmann, D. C1 - 53714 C2 - 44922 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland SP - 1539 TI - BCL10-Bridging CARDs to immune activation. JO - Front. Immunol. VL - 9 IS - JUL PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - [This corrects the article DOI: 10.3389/fimmu.2018.00979.]. AU - Ghosh, S.* AU - Drexler, I.* AU - Bhatia, S.* AU - Adler, H. AU - Gennery, A.R.* AU - Borkhardt, A.* C1 - 54488 C2 - 45564 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland SP - 2197 TI - Erratum: Corrigendum: Interleukin-2-inducible T-cell kinase deficiency-new patients, new insight? (Frontiers in immunology (2018) 9 (979)). JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Patients with primary immunodeficiency can be prone to severe Epstein-Barr virus (EBV) associated immune dysregulation. Individuals with mutations in the interleukin-2-inducible T-cell kinase () gene experience Hodgkin and non-Hodgkin lymphoma, EBV lymphoproliferative disease, hemophagocytic lymphohistiocytosis, and dysgammaglobulinemia. In this review, we give an update on further reported patients. We believe that current clinical data advocate early definitive treatment by hematopoietic stem cell transplantation, as transplant outcome in primary immunodeficiency disorders in general has gradually improved in recent years. Furthermore, we summarize experimental data in the murine model to provide further insight of pathophysiology in ITK deficiency. AU - Ghosh, S.* AU - Drexler, I.* AU - Bhatia, S.* AU - Adler, H. AU - Gennery, A.R.* AU - Borkhardt, A.* C1 - 54604 C2 - 45688 TI - Interleukin-2-inducible T-cell kinase deficiency-new patients, new insight? JO - Front. Immunol. VL - 9 PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - A novel vaccine against bovine viral diarrhea (BVD) induced pathogenic antibody production in 5-10% of BVD-vaccinated cows. Transfer of these antibodies via colostrum caused Bovine neonatal pancytopenia (BNP) in calves, with a lethality rate of 90%. The exact immunological mechanisms behind the onset of BNP are not fully understood to date. To gain further insight into these mechanisms, we analyzed the immune proteome from alloreactive antibody producers (BNP cows) and non-responders. After stimulation of peripheral blood derived lymphocytes (PBL), we detected distinctly deviant expression levels of several master regulators of immune responses in BNP cells, pointing to a changed immune phenotype with severe dysregulation of immune response in BNP cows. Interestingly, we also found this response pattern in 22% of non-BVD-vaccinated cows, indicating a genetic predisposition of this immune deviant (ID) phenotype in cattle. We additionally analyzed the functional correlation of the ID phenotype with 10 health parameters and 6 diseases in a retrospective study over 38 months. The significantly increased prevalence of mastitis among ID cows emphasizes the clinical relevance of this deviant immune response and its potential impact on the ability to fight infections. AU - Lutterberg, K.* AU - Kleinwort, K.J.H.* AU - Hobmaier, B.F.* AU - Hauck, S.M. AU - Nuske, S.* AU - Scholz, A.M.* AU - Deeg, C.A.* C1 - 55008 C2 - 46011 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - A functionally different immune phenotype in cattle is associated with higher mastitis incidence. JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - The envelope of Human Immunodeficiency Virus type 1 (HIV-1) consists of a liquid-ordered membrane enriched in raft lipids and containing the viral glycoproteins. Previous studies demonstrated that changes in viral membrane lipid composition affecting membrane structure or curvature can impair infectivity. Here, we describe novel antiviral compounds that were identified by screening compound libraries based on raft lipid-like scaffolds. Three distinct molecular structures were chosen for mode-of-action studies, a sterol derivative (J391B), a sphingosine derivative (J582C) and a long aliphatic chain derivative (IBS70). All three target the viral membrane and inhibit virus infectivity at the stage of fusion without perturbing virus stability or affecting virion-associated envelope glycoproteins. Their effect did not depend on the expressed envelope glycoproteins or a specific entry route, being equally strong in HIV pseudotypes carrying VSV-G or MLV-Env glycoproteins. Labeling with laurdan, a reporter of membrane order, revealed different membrane structure alterations upon compound treatment of HIV-1, which correlated with loss of infectivity. J582C and IBS70 decreased membrane order in distinctive ways, whereas J391B increased membrane order. The compounds' effects on membrane order were reproduced in liposomes generated from extracted HIV lipids and thus independent both of virion proteins and of membrane leaflet asymmetry. Remarkably, increase of membrane order by J391B required phosphatidylserine, a lipid enriched in the HIV envelope. Counterintuitively, mixtures of two compounds with opposite effects on membrane order, J582C and J391B, did not neutralize each other but synergistically inhibited HIV infection. Thus, altering membrane order, which can occur by different mechanisms, constitutes a novel antiviral mode of action that may be of general relevance for enveloped viruses and difficult to overcome by resistance development. AU - Nieto-Garai, J.A.* AU - Glass, B.* AU - Bunn, C.* AU - Giese, M.* AU - Jennings, G.* AU - Brankatschk, B. AU - Agarwal, S.* AU - Boerner, K.* AU - Contreras, F.X.* AU - Knoelker, H.* AU - Zankl, C.* AU - Simons, K.* AU - Schroeder, C.* AU - Lorizate, M.* AU - Kraeusslich, H.* C1 - 54266 C2 - 45384 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland TI - Lipidomimetic compounds act as HIV-1 entry inhibitors by altering viral membrane structure. JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Assembly of the CARD11/CARMA1-BCL10-MALT1 (CBM) signaling complex upon T or B cell antigen receptor (TCR or BCR) engagement drives lymphocyte activation. Recruitment of pre-assembled BCL10-MALT1 complexes to CARD11 fosters activation of the MALT1 protease and canonical NF-B signaling. Structural data and assays have suggested that CARD11 acts as a seed that nucleates the assembly of BCL10 filaments, but the relevance of these findings for CBM complex assembly in cells remains unresolved. To uncouple cellular CARD11 recruitment of BCL10 and BCL10 filament assembly, we generated a BCL10-CARD11 fusion protein that links the C-terminus of BCL10 to the N-terminus of CARD11. When stably expressed in CARD11 KO Jurkat T cells, the BCL10-CARD11 fusion induced constitutive MALT1 activation. Furthermore, in CARD11 KO BJAB B cells, BCL10-CARD11 promoted constitutive NF-B activation to a similar extent as CARD11 containing oncogenic driver mutations. Using structure-guided destructive mutations in the CARD11-BCL10 (CARD11 R35A) or BCL10-BCL10 (BCL10 R42E) interfaces, we demonstrate that chronic activation by the BCL10-CARD11 fusion protein was independent of the CARD11 CARD. However, activation strictly relied upon the ability of the BCL10 CARD to form oligomers. Thus, by combining distinct CARD mutations in the context of constitutively active BCL10-CARD11 fusion proteins, we provide evidence that BCL10-MALT1 recruitment to CARD11 and BCL10 oligomerization are interconnected processes, which bridge the CARD11 seed to downstream pathways in lymphocytes. AU - Seeholzer, T. AU - Kurz, S. AU - Schlauderer, F.* AU - Woods, S. AU - Gehring, T. AU - Widmann, S. AU - Lammens, K.* AU - Krappmann, D. C1 - 54823 C2 - 45846 CY - Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland SP - 2695 TI - BCL10-CARD11 fusion mimics an active CARD11 seed that triggers constitutive BCL10 oligomerization and lymphocyte activation. JO - Front. Immunol. VL - 9 IS - NOV PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - T follicular helper (TFH) cells are an integral part of humoral immunity by providing help to B cells to produce high-affinity antibodies. The TFH precursor compartment circulates in the blood and TFH cell dysregulation is implied in various autoimmune diseases including type 1 diabetes (T1D). Symptomatic T1D is preceded by a preclinical phase (indicated by the presence of islet autoantibodies) with a highly variable progression time to the symptomatic disease. This heterogeneity points toward differences in immune activation in children with a fast versus slow progressor phenotype. In the context of T1D, previous studies on TFH cells have mainly focused on the clinically active state of the disease. In this review article, we aim to specifically discuss recent insights on TFH cells in human islet autoimmunity before the onset of symptomatic T1D. Furthermore, we will highlight advances in the field of TFH differentiation and function during human islet autoimmunity. Specifically, we will focus on the regulation of TFH cells by microRNAs (miRNAs), as well as on the potential use of miRNAs as biomarkers to predict disease progression time and as future drug targets to interfere with autoimmune activation. AU - Serr, I. AU - Daniel, C. C1 - 54037 C2 - 45226 TI - Regulation of T follicular helper cells in islet autoimmunity. JO - Front. Immunol. VL - 9 PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Immunoglobulin G (IgG), a glycoprotein secreted by plasma B-cells, plays a major role in the human adaptive immune response and are associated with a wide range of diseases. Glycosylation of the Fc binding region of IgGs, responsible for the antibody's effector function, is essential for prompting a proper immune response. This study focuses on the general genetic impact on IgG glycosylation as well as corresponding subclass specificities. To identify genetic loci involved in IgG glycosylation, we performed a genome-wide association study (GWAS) on liquid chromatography electrospray mass spectrometry (LC-ESI-MS)-measured IgG glycopeptides of 1,823 individuals in the Cooperative Health Research in the Augsburg Region (KORA F4) study cohort. In addition, we performed GWAS on subclass-specific ratios of IgG glycans to gain power in identifying genetic factors underlying single enzymatic steps in the glycosylation pathways. We replicated our findings in 1,836 individuals from the Leiden Longevity Study (LLS). We were able to show subclass-specific genetic influences on single IgG glycan structures. The replicated results indicate that, in addition to genes encoding for glycosyltransferases (i.e.,, and), other genetic loci have strong influences on the IgG glycosylation patterns. A novel locus on chromosome 1, harboring, which encodes for a transcription factor of the runt domain-containing family, is associated with decreased galactosylation. Interestingly, members of thefamily are cross-regulated, andis involved in both IgA class switching and B-cell maturation as well as T-cell differentiation and apoptosis. Besides the involvement of glycosyltransferases in IgG glycosylation, we suggest that, due to the impact of variants within, potentially mechanisms involved in B-cell activation and T-cell differentiation during the immune response as well as cell migration and invasion involve IgG glycosylation. AU - Wahl, A. AU - van den Akker, E.* AU - Klaric, L.* AU - Štambuk, J.* AU - Benedetti, E. AU - Plomp, R.* AU - Razdorov, G.* AU - Trbojević-Akmačić, I.* AU - Deelen, J.* AU - van Heemst, D.* AU - Eline Slagboom, P.* AU - Vučković, F.* AU - Grallert, H. AU - Krumsiek, J. AU - Strauch, K. AU - Peters, A. AU - Meitinger, T. AU - Hayward, C.* AU - Wuhrer, M.* AU - Beekman, M.* AU - Lauc, G.* AU - Gieger, C. C1 - 53092 C2 - 44448 CY - Lausanne TI - Genome-wide association study on immunoglobulin G glycosylation patterns. JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Many current vaccines are less immunogenic and less effective in elderly compared to younger adults due to age-related changes of the immune system. Most vaccines utilized in the elderly contain antigens, which the target population has had previous contact with due to previous vaccination or infection. Therefore, most studies investigating vaccine-induced immune responses in the elderly do not analyze responses to neo-antigens but rather booster responses. However, age-related differences in the immune response could differentially affect primary versus recall responses. We therefore investigated the impact of age on primary and recall antibody responses following hepatitis B vaccination in young and older adults. Focused gene expression profiling was performed before and 1 day after the vaccination in order to identify gene signatures predicting antibody responses. Young (20-40 years;  = 24) and elderly (>60 years;  = 17) healthy volunteers received either a primary series (no prior vaccination) or a single booster shot (documented primary vaccination more than 10 years ago). Antibody titers were determined at days 0, 7, and 28, as well as 6 months after the vaccination. After primary vaccination, antibody responses were lower and delayed in the elderly compared to young adults. Non-responders after the three-dose primary series were only observed in the elderly group. Maximum antibody concentrations after booster vaccination were similar in both age groups. Focused gene expression profiling identified 29 transcripts that correlated with age at baseline and clustered in a network centered around type I interferons and pro-inflammatory cytokines. In addition, smaller 8- and 6-gene signatures were identified at baseline that associated with vaccine responsiveness during primary and booster vaccination, respectively. When evaluating the kinetic changes in gene expression profiles before and after primary vaccination, a 33-gene signature, dominated by IFN-signaling, pro-inflammatory cytokines, inflammasome components, and immune cell subset markers, was uncovered that was associated with vaccine responsiveness. By contrast, no such transcripts were identified during booster vaccination. Our results document that primary differs from booster vaccination in old age, in regard to antibody responses as well as at the level of gene signatures. Clinical Trial Registration: www.clinicaltrialsregister.eu, this trial was registered at the EU Clinical Trial Register (EU-CTR) with the EUDRACT-Nr. 2013-002589-38. AU - Weinberger, B.* AU - Haks, M.C.* AU - de Paus, R.A.* AU - Ottenhoff, T.H.M.* AU - Bauer, T. AU - Grubeck-Loebenstein, B.* C1 - 53587 C2 - 44695 CY - Lausanne TI - Impaired immune response to primary but not to booster vaccination against hepatitis B in older adults. JO - Front. Immunol. VL - 9 PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Newcastle disease (ND), caused by infections with virulent strains of Newcastle disease virus (NDV), is one of the most important infectious disease affecting wild, peridomestic, and domestic birds worldwide. Vaccines constructed from live, low-virulence (lentogenic) viruses are the most accepted prevention and control strategies for combating ND in poultry across the globe. Avian macrophages are one of the first cell lines of defense against microbial infection, responding to signals in the microenvironment. Although macrophages are considered to be one of the main target cells for NDV infection in vivo, very little is known about the ability of NDV to infect chicken macrophages, and virulence mechanisms of NDV as well as the polarized activation patterns of macrophages and correlation with viral infection and replication. In the present study, a cell culture model (chicken bone marrow macrophage cell line HD11) and three different virulence and genotypes of NDV (including class II virulent NA-1, class II lentogenic LaSota, and class I lentogenic F55) were used to solve the above underlying questions. Our data indicated that all three NDV strains had similar replication rates during the early stages of infection. Virulent NDV titers were shown to increase compared to the other lentogenic strains, and this growth was associated with a strong upregulation of both pro-inflammatory M1-like markers/cytokines and anti-inflammatory M2-like markers/cytokines in chicken macrophages. Virulent NDV was found to block toll-like receptor (TLR) 7 expression, inducing higher expression of type I interferons in chicken macrophages at the late stage of viral infection. Only virulent NDV replication can be inhibited by pretreatment with TLR7 ligand. Overall, this study demonstrated that virulent NDV activates a M1-/M2-like mixed polarized activation of chicken macrophages by inhibition of TLR7, resulting in enhanced replication compared to lentogenic viruses. AU - Zhang, P.* AU - Ding, Z.* AU - Liu, X.* AU - Chen, Y.* AU - Li, J.* AU - Tao, Z.* AU - Fei, Y.* AU - Xue, C.* AU - Qian, J.* AU - Wang, X.* AU - Li, Q.* AU - Stöger, T. AU - Chen, J.* AU - Bi, Y.* AU - Yin, R.* C1 - 53421 C2 - 44698 CY - Lausanne TI - Enhanced replication of virulent Newcastle disease virus in chicken macrophages is due to polarized activation of cells by inhibition of TLR7. JO - Front. Immunol. VL - 9 IS - APR PB - Frontiers Media Sa PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disease, characterized by damage of lung epithelial cells, excessive deposition of extracellular matrix in the lung interstitium, and enhanced activation and proliferation of fibroblasts. S100a4, also termed FSP-1 (fibroblast-specific protein-1), was previously considered as a marker of fibroblasts but recent findings in renal and liver fibrosis indicated that M2 macrophages are an important cellular source of S100a4. Thus, we hypothesized that also in pulmonary fibrosis, M2 macrophages produce and secrete S100a4, and that secreted S100a4 induces the proliferation and activation of fibroblasts. To prove this hypothesis, we comprehensively characterized two established mouse models of lung fibrosis: infection of IFN-γR-/-mice with MHV-68 and intratracheal application of bleomycin to C57BL/6 mice. We further provide in vitro data using primary macrophages and fibroblasts to investigate the mechanism by which S100A4 exerts its effects. Finally, we inhibit S100a4 in vivo in the bleomycin-induced lung fibrosis model by treatment with niclosamide. Our data suggest that S100a4 is produced and secreted by M2 polarized alveolar macrophages and enhances the proliferation and activation of lung fibroblasts. Inhibition of S100a4 might represent a potential therapeutic strategy for pulmonary fibrosis. AU - Zhang, W. AU - Ohno, S. AU - Steer, B. AU - Klee, S. AU - Staab-Weijnitz, C.A. AU - Wagner, D.E. AU - Lehmann, M. AU - Stöger, T. AU - Königshoff, M. AU - Adler, H. C1 - 53610 C2 - 44708 TI - S100a4 is secreted by alternatively activated alveolar macrophages and promotes activation of lung fibroblasts in pulmonary fibrosis. JO - Front. Immunol. VL - 9 IS - JUN PY - 2018 SN - 1664-3224 ER - TY - JOUR AB - Short-chain fatty acids (SCFAs), which are generated by the bacterial fermentation of dietary fibers, promote expansion of regulatory T cells (Tregs). Potential therapeutic value of SCFAs has been recently highlighted in the experimental models of T cell-mediated autoimmunity and allergic inflammation. These studies suggest that physiological intestinal concentrations of SCFAs within the millimolar range are crucial for dampening inflammation-mediated processes. Here, we describe opposing effects of SCFAs on T cell-mediated immune responses. In accordance with published data, lower butyrate concentrations facilitated differentiation of Tregs in vitro and in vivo under steady-state conditions. In contrast, higher concentrations of butyrate induced expression of the transcription factor T-bet in all investigated T cell subsets resulting in IFN-γ-producing Tregs or conventional T cells. This effect was mediated by the inhibition of histone deacetylase activity and was independent of SCFA-receptors FFA2 and FFA3 as well as of Na + - coupled SCFA transporter Slc5a8. Importantly, while butyrate was not able to induce the generation of Tregs in the absence of TGF-β1, the expression of T-bet and IFN-γ was triggered upon stimulation of CD4 + T cells with this SCFA alone. Moreover, the treatment of germ-free mice with butyrate enhanced the expression of T-bet and IFN-γ during acute colitis. Our data reveal that, depending on its concentration and immunological milieu, butyrate may exert either beneficial or detrimental effects on the mucosal immune system. AU - Kespohl, M.* AU - Vachharajani, N.* AU - Luu, M.* AU - Harb, H.* AU - Pautz, S.* AU - Wolff, S.* AU - Sillner, N. AU - Walker, A. AU - Schmitt-Kopplin, P. AU - Boettger, T.* AU - Renz, H.* AU - Offermanns, S.* AU - Steinhoff, U.* AU - Visekruna, A.* C1 - 51794 C2 - 43476 CY - Lausanne TI - The microbial metabolite butyrate induces expression of Th1- associated factors in cD4+ T cells. JO - Front. Immunol. VL - 8 PB - Frontiers Media Sa PY - 2017 SN - 1664-3224 ER - TY - JOUR AB - Obesity-related adipose tissue (AT) inflammation that promotes metabolic dysregulation is associated with increased AT mast cell numbers. Mast cells are potent inducers of inflammatory responses and could potentially contribute to obesity-induced AT inflammation and metabolic dysregulation. Conflicting findings were reported on obesity-related metabolic dysfunction in mast cell-deficient mice, thus creating a controversy that has not been resolved to date. Whereas traditional Kit hypomorphic mast cell-deficient strains featured reduced diet-induced obesity and diabetes, a Kit-independent model of mast cell deficiency, Cpa3(Cre/+) mice, displayed no alterations in obesity and insulin sensitivity. Herein, we analyzed diet-induced obesity in Mcpt5-Cre R-DTA mice, in which the lack of mast cells is caused by a principle different from mast cell deficiency in Cpa3(Cre/+) mice or Kit mutations. We observed no difference between mast cell-deficient and -proficient mice in diet-induced obesity with regards to weight gain, glucose tolerance, insulin resistance, metabolic parameters, hepatic steatosis, and AT or liver inflammation. We conclude that mast cells play no essential role in obesity and related pathologies. AU - Chmelar, J.* AU - Chatzigeorgiou, A.* AU - Chung, K.-J. AU - Prucnal, M.* AU - Voehringer, D.* AU - Roers, A.* AU - Chavakis, T. C1 - 50112 C2 - 42045 CY - Lausanne TI - No role for mast cells in obesity-related metabolic dysregulation. JO - Front. Immunol. VL - 7 PB - Frontiers Media Sa PY - 2016 SN - 1664-3224 ER - TY - JOUR AB - The human complement factor H-related protein-3 (FHR-3) is a soluble regulator of the complement system. Homozygous cfhr3/1 deletion is a genetic risk factor for the autoimmune form of atypical hemolytic-uremic syndrome (aHUS), while also found to be protective in age-related macular degeneration (AMD). The precise function of FHR-3 remains to be fully characterized. We generated four mouse monoclonal antibodies (mAbs) for FHR-3 (RETC) without cross-reactivity to the complement factor H (FH)-family. These antibodies detected FHR-3 from human serum with a mean concentration of 1 μg/mL. FHR-3 levels in patients were significantly increased in sera from systemic lupus erythematosus, rheumatoid arthritis, and polymyalgia rheumatica but remained almost unchanged in samples from AMD or aHUS patients. Moreover, by immunostaining of an aged human donor retina, we discovered a local FHR-3 production by microglia/macrophages. The mAb RETC-2 modulated FHR-3 binding to C3b but not the binding of FHR-3 to heparin. Interestingly, FHR-3 competed with FH for binding C3b and the mAb RETC-2 reduced the interaction of FHR-3 and C3b, resulting in increased FH binding. Our results unveil a previously unknown systemic involvement of FHR-3 in rheumatoid diseases and a putative local role of FHR-3 mediated by microglia/macrophages in the damaged retina. We conclude that the local FHR-3/FH equilibrium in AMD is a potential therapeutic target, which can be modulated by our specific mAb RETC-2. AU - Schaefer, N.* AU - Grosche, A.* AU - Reinders, J.* AU - Hauck, S.M. AU - Pouw, R.B.* AU - Kuijpers, T.W.* AU - Wouters, D.* AU - Ehrenstein, B.* AU - Enzmann, V.* AU - Zipfel, P.F.* AU - Skerka, C.* AU - Pauly, D.* C1 - 50111 C2 - 42044 CY - Lausanne SP - 542 TI - Complement regulator FHR-3 is elevated either locally or systemically in a selection of autoimmune diseases. JO - Front. Immunol. VL - 7 PB - Frontiers Media Sa PY - 2016 SN - 1664-3224 ER - TY - JOUR AB - Natural killer (NK) cells play a pivotal role in the first line of defense against cancer. NK cells that are deficient in CD3 and a clonal T cell receptor (TCR) can be subdivided into two major subtypes, CD56dimCD16+ cytotoxic and CD56brightCD16− immunoregulatory NK cells. Cytotoxic NK cells not only directly kill tumor cells without previous stimulation by cytotoxic effector molecules, such as perforin and granzymes or via death receptor interactions, but also act as regulatory cells for the immune system by secreting cytokines and chemokines. The aim of this review is to highlight therapeutic strategies utilizing autologous and allogenic NK cells, combinations of NK cells with monoclonal antibodies to induce antibody-dependent cellular cytotoxicity, or immune checkpoint inhibitors. Additionally, we discuss the use of chimeric antigen receptor-engineered NK cells in cancer immunotherapy. AU - Shevtsov, M* AU - Multhoff, G. C1 - 50011 C2 - 41967 CY - Lausanne TI - Immunological and translational aspects of NK cell-based antitumor immunotherapies. JO - Front. Immunol. VL - 7 PB - Frontiers Media Sa PY - 2016 SN - 1664-3224 ER - TY - JOUR AB - T-cell responses to the immediate-early 1 (IE-1) protein of human cytomegalovirus (HCMV) are associated with protection from viral disease. Thus, IE-1 is a promising target for immunotherapy. CD8 T-cell responses to IE-1 are generally strong. In contrast, CD4 T-cell responses to IE-1 were described to be comparatively infrequent or undetectable in HCMV carriers, and information on their target epitopes and their function has been limited. To analyze the repertoire of IE-1-specific CD4 T cells, we expanded them from healthy donors with autologous IE-1-expressing mini-Epstein-Barr virus-transformed B-cell lines and established IE-1-specific CD4 T-cell clones. Clones from seven out of seven HCMV-positive donors recognized endogenously processed IE-1 epitopes restricted through HLA-DR, DQ, or DP. Three to seven IE-1 epitopes were recognized per donor. Cumulatively, about 27 different HLA/peptide class II complexes were recognized by 117 IE-1-specific clones. Our results suggest that a highly diversified repertoire of IE-1-specific CD4 T cells targeting multiple epitopes is usually present in healthy HCMV carriers. Therefore, multiepitope approaches to immunomonitoring and immunotherapy will make optimal use of this potentially important class of HCMV-specific effector cells. AU - Ameres, S. AU - Liang, X. AU - Wiesner, M. AU - Mautner, J. AU - Moosmann, A. C1 - 47499 C2 - 40605 TI - A diverse repertoire of CD4 T cells targets the immediate-early 1 protein of human cytomegalovirus. JO - Front. Immunol. VL - 6 PY - 2015 SN - 1664-3224 ER - TY - JOUR AB - Heat-shock protein 70 (Hsp70) is frequently found on the plasma membrane of a large number of malignant tumors including non-small cell lung cancer (NSCLC) and gets released into the blood circulation in lipid vesicles. On the one hand, a membrane (m)Hsp70-positive phenotype correlates with a high aggressiveness of the tumor; on the other hand, mHsp70 serves as a target for natural killer (NK) cells that had been pre-stimulated with Hsp70-peptide TKD plus low-dose interleukin-2 (TKD/IL-2). Following activation, NK cells show an up-regulated expression of activatory C-type lectin receptors, such as CD94/NKG2C, NKG2D, and natural cytotoxicity receptors (NCRs; NKp44, NKp46, and NKp30) and thereby gain the capacity to kill mHsp70-positive tumor cells. With respect to these results, the efficacy of ex vivo TKD/IL-2 stimulated, autologous NK cells is currently tested in a proof-of-concept phase II clinical trial in patients with squamous cell NSCLC after radiochemotherapy (RCT) at the TUM. Inclusion criteria are histological proven, non-resectable NSCLC in stage IIIA/IIIB, clinical responses to RCT and a mHsp70-positive tumor phenotype. The mHsp70 status is determined in the serum of patients using the lipHsp70 ELISA test, which enables the quantification of liposomal and free Hsp70. Squamous cell and adeno NSCLC patients had significantly higher serum Hsp70 levels than healthy controls. A significant correlation of serum Hsp70 levels with the gross tumor volume was shown for adeno and squamous cell NSCLC. However, significantly elevated ratios of activated CD69(+)/CD94(+) NK cells that are associated with low serum Hsp70 levels were observed only in patients with squamous cell lung cancer. These data might provide a first hint that squamous cell NSCLC is more immunogenic than adeno NSCLC. AU - Gunther, S.* AU - Ostheimer, C.* AU - Stangl, S.* AU - Specht, H.M.* AU - Mozes, P.* AU - Jesinghaus, M.* AU - Vordermark, D.* AU - Combs, S.E. AU - Peltz, F.* AU - Jung, M.P.* AU - Multhoff, G. C1 - 47338 C2 - 39311 TI - Correlation of Hsp70 serum levels with gross tumor volume and composition of lmphocyte subpopulations in patients with squamous cell and adeno non-small cell lung cancer. JO - Front. Immunol. VL - 6 PY - 2015 SN - 1664-3224 ER - TY - JOUR AB - Heat shock protein 70 (Hsp70) is frequently overexpressed in tumor cells. An unusual cell surface localization could be demonstrated on a large variety of solid tumors including lung, colorectal, breast, squamous cell carcinomas of the head and neck, prostate and pancreatic carcinomas, glioblastomas, sarcomas and hematological malignancies, but not on corresponding normal tissues. A membrane (m)Hsp70-positive phenotype can be determined either directly on single cell suspensions of tumor biopsies by flow cytometry using cmHsp70.1 monoclonal antibody or indirectly in the serum of patients using a novel lipHsp70 ELISA. A mHsp70-positive tumor phenotype has been associated with highly aggressive tumors, causing invasion and metastases and resistance to cell death. However, natural killer (NK), but not T cells were found to kill mHsp70-positive tumor cells after activation with a naturally occurring Hsp70 peptide (TKD) plus low dose IL-2 (TKD/IL-2). Safety and tolerability of ex vivo TKD/IL-2 stimulated, autologous NK cells has been demonstrated in patients with metastasized colorectal and NSCLC in a phase I clinical trial. Based on promising clinical results of the previous study, a phase II randomized clinical study was initiated in 2015. The primary objective of this multicenter proof-of-concept trial is to examine whether an adjuvant treatment of NSCLC patients after platinum based radiochemotherapy with TKD/IL-2 activated, autologous NK cells is clinically effective. As a mHsp70-positive tumor phenotype is associated with poor clinical outcome only mHsp70-positive tumor patients will be recruited into the trial. The primary endpoint of this study will be the comparison of the progression-free survival of patients treated with ex vivo activated NK cells compared to patients who were treated with radiochemotherapy alone. As secondary endpoints overall survival, toxicity, quality-of-life and biological responses will be determined in both study groups. AU - Specht, H.M.* AU - Ahrens, N.* AU - Blankenstein, C.* AU - Duell, T.H.G.* AU - Fietkau, R.J.* AU - Gaipl, U.S.* AU - Günther, C.* AU - Gunther, S.* AU - Habl, G.* AU - Hautmann, H.* AU - Hautmann, M.G.* AU - Huber, R.M.* AU - Molls, M.M.* AU - Offner, R.* AU - Rödel, C.M.* AU - Rödel, F.* AU - Schuetz, M.* AU - Combs, S.E.* AU - Multhoff, G. C1 - 44427 C2 - 36917 TI - Heat shock protein 70 (Hsp70) peptide activated Natural Killer (NK) cells for the treatment of patients with Non-Small Cell Lung Cancer (NSCLC) after radiochemotherapy (RCTx) - from preclinical studies to a clinical phase II trial. JO - Front. Immunol. VL - 6 PY - 2015 SN - 1664-3224 ER - TY - JOUR AB - In contrast to the past reliance on morphology, the identification and enumeration of blood monocytes are nowadays done with monoclonal antibodies and flow cytometry and this allows for subdivision into classical, intermediate, and non-classical monocytes. Using specific cell surface markers, dendritic cells in blood can be segregated from these monocytes. While in the past, changes in monocyte numbers as determined in standard hematology counters have not had any relevant clinical impact, the subset analysis now has uncovered informative changes that may be used in management of disease. AU - Ziegler-Heitbrock, L. C1 - 46761 C2 - 37788 CY - Lausanne TI - Blood monocytes and their subsets: Established features and open questions. JO - Front. Immunol. VL - 6 PB - Frontiers Media Sa PY - 2015 SN - 1664-3224 ER - TY - JOUR AB - Members of the heat shock protein 70 (HSP70) family play an important role in assisting protein folding, preventing protein aggregation and transport of proteins across membranes under physiological conditions. Following environmental (i.e., irradiation, chemotherapy), physiological (i.e., cell growth, differentiation), and pathophysiological (i.e., inflammation, tumorigenesis) stress, the synthesis of heat shock proteins (HSPs) is highly up-regulated, whereas protein synthesis in general is reduced. In contrast to normal cells, many tumor entities including hepatocellular carcinoma (HCC) overexpress HSP70, the major-stress-inducible member of the HSP70 family, present it on their cell surface and secrete it into the extracellular milieu. Herein, the prognostic relevance of serum HSP70 levels in patients with chronic hepatitis (CH; n = 50), liver cirrhosis (LC; n = 46), and HCC (n = 47) was analyzed. Similar to other tumor entities, HSP70 is also present on the surface of primary HCC cells. The staining intensity of intracellular HSP70 in HCC tissue is stronger compared to control and cirrhotic liver sections. HSP70 serum levels in all HCC patients were significantly higher compared to a control group without liver disease (n = 40). No significant age- and gender-related differences in HSP70 serum levels were observed in male and female healthy human volunteers (n = 86). Patients with CH (n = 50) revealed significantly higher HSP70 serum levels compared to the control group, however, these values were significantly lower than those of HCC patients (n = 47). Furthermore, a subgroup of patients with LC who subsequently developed HCC (LC-HCC, n = 13) revealed higher HSP70 serum levels than patients with LC (n = 46, p = 0.05). These data indicate that serum HSP70 levels are consecutively increased in patients with CH, LC and liver carcinomas and thus might have a prognostic value. AU - Gehrmann, M.* AU - Cervello, M.* AU - Montalto, G.* AU - Cappello, F.* AU - Gulino, A.* AU - Knape, C.* AU - Specht, H.M.* AU - Multhoff, G. C1 - 31847 C2 - 34813 CY - Lausanne TI - Heat shock protein 70 serum levels differ significantly in patients with chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. JO - Front. Immunol. VL - 5 PB - Frontiers Research Foundation PY - 2014 SN - 1664-3224 ER - TY - JOUR AB - In Western Europe, Hymenoptera venom allergy (HVA) primarily relates to venoms of the honeybee and the common yellow jacket. In contrast to other allergen sources, only a few major components of Hymenoptera venoms had been characterized until recently. Improved expression systems and proteomic detection strategies have allowed the identification and characterization of a wide range of additional allergens. The field of HVA research has moved rapidly from focusing on venom extract and single major allergens to a molecular understanding of the entire "venome" as a system of unique and characteristic components. An increasing number of such components has been identified, characterized regarding function, and assessed for allergenic potential. Moreover, advanced expression strategies for recombinant production of venom allergens allow selective modification of molecules and provide insight into different types of immunoglobulin E reactivities and sensitization patterns. The obtained information contributes to an increased diagnostic precision in HVA and may serve for monitoring, re-evaluation, and improvement of current therapeutic strategies. AU - Spillner, E.* AU - Blank, S. AU - Jakob, T.* C1 - 30874 C2 - 33983 TI - Hymenoptera allergens: From venom to "venome". JO - Front. Immunol. VL - 5 PY - 2014 SN - 1664-3224 ER - TY - JOUR AB - CD8(+) T cell immune responses provide immediate protection against primary infection and durable memory capable of rapidly fighting off re-infection. Immediate protection and lasting memory are implemented by phenotypically and functionally distinct T cell subsets. While it is now widely accepted that these diverge from a common source of naïve T cells (T(n)), the developmental relation and succession of effector and memory T cell subsets is still under intense debate. Recently, a distinct memory T cell subset has been suggested to possess stem cell-like features, sparking the hope to harness its capacity for self-renewal and diversification for successful therapy of chronic infections or malignant diseases. In this review we highlight current developmental models of memory generation, T cell subset diversification and T cell stemness. We discuss the importance of single cell monitoring techniques for adequately mapping these developmental processes and take a brief look at signaling components active in the putative stem cell-like memory T cell compartment. AU - Buchholz, V.R.* AU - Gräf, P.* AU - Busch, D.H. C1 - 28007 C2 - 32899 TI - The smallest unit: Effector and memory CD8+ T cell differentiation on the single cell level. JO - Front. Immunol. VL - 4 PB - Frontiers PY - 2013 SN - 1664-3224 ER - TY - JOUR AB - In a nomenclature proposal published in 2010 monocytes were subdivided into classical and non-classical cells and in addition an intermediate monocyte subset was proposed. Over the last couple of years many studies have analyzed these intermediate cells, their characteristics have been described, and their expansion has been documented in many clinical settings. While these cells appear to be in transition from classical to non-classical monocytes and hence may not form a distinct cell population in a strict sense, their separate analysis and enumeration is warranted in health and disease. AU - Ziegler-Heitbrock, L. AU - Hofer, T.P. C1 - 27515 C2 - 32708 TI - Toward a refined definition of monocyte subsets. JO - Front. Immunol. VL - 4 PB - Frontiers Research Foundation PY - 2013 SN - 1664-3224 ER - TY - JOUR AB - Chronic inflammatory mediators exert pleiotropic effects in the development of cancer. On the one hand, inflammation favors carcinogenesis, malignant transformation, tumor growth, invasion, and metastatic spread; on the other hand inflammation can stimulate immune effector mechanisms that might limit tumor growth. The link between cancer and inflammation depends on intrinsic and extrinsic pathways. Both pathways result in the activation of transcription factors such as NF-κB, STAT-3, and HIF-1 and in accumulation of tumorigenic factors in tumor and microenvironment. STAT-3 and NF-κB interact at multiple levels and thereby boost tumor-associated inflammation which can suppress anti-tumor immune responses. These factors also promote tumor growth, progression, and metastatic spread. IL-1, IL-6, TNF, and PGHS-2 are key mediators of an inflammatory milieu by modulating the expression of tumor-promoting factors. In this review we concentrate on the crucial role of pro-inflammatory mediators in inflammation-driven carcinogenesis and outline molecular mechanisms of IL-1 signaling in tumors. In addition, we elucidate the dual roles of stress proteins as danger signals in the development of anti-cancer immunity and anti-apoptotic functions. AU - Multhoff, G. AU - Molls, M.* AU - Radons, J.* C1 - 7160 C2 - 29500 TI - Chronic inflammation in cancer development. JO - Front. Immunol. VL - 2 IS - JAN PB - Frontiers Media S.A. PY - 2012 SN - 1664-3224 ER -