TY  - JOUR
AB  - Resting conventional T cells (Tconv) can be distinguished from T regulatory cells (Treg) by the canonical markers FOXP3, CD25 and CD127. However, the expression of these proteins alters after T-cell activation leading to overlap between Tconv and Treg. The objective of this study was to distinguish resting and antigen-responsive T effector (Tconv) and Treg using single-cell technologies. CD4+ Treg and Tconv cells were stimulated with antigen and responsive and non-responsive populations processed for targeted and non-targeted single-cell RNAseq. Machine learning was used to generate a limited set of genes that could distinguish responding and non-responding Treg and Tconv cells and which was used for single-cell multiplex qPCR and to design a flow cytometry panel. Targeted scRNAseq clearly distinguished the four-cell populations. A minimal set of 27 genes was identified by machine learning algorithms to provide discrimination of the four populations at >95% accuracy. In all, 15 of the genes were validated to be differentially expressed by single-cell multiplex qPCR. Discrimination of responding Treg from responding Tconv could be achieved by a flow cytometry strategy that included staining for CD25, CD127, FOXP3, IKZF2, ITGA4, and the novel marker TRIM which was strongly expressed in Tconv and weakly expressed in both responding and non-responding Treg. A minimal set of genes was identified that discriminates responding and non-responding CD4+ Treg and Tconv cells and, which have identified TRIM as a marker to distinguish Treg by flow cytometry.
AU  - Reinhardt, J.*
AU  - Sharma, V.
AU  - Stavridou, A.*
AU  - Lindner, A.
AU  - Reinhardt, S.*
AU  - Petzold, A.*
AU  - Lesche, M.*
AU  - Rost, F.*
AU  - Bonifacio, E.
AU  - Eugster, A.*
C1  - 64496
C2  - 52230
SP  - 121-137
TI  - Distinguishing activated T regulatory cell and T conventional cells by single-cell technologies.
JO  - Immunology
VL  - 166
IS  - 1
PY  - 2022
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - T-helper cell 17 (Th17) mediated inflammation is associated with various diseases including autoimmune encephalitis, inflammatory bowel disease and lung diseases such as chronic obstructive pulmonary disease and asthma. Differentiation into distinct Th subtypes needs to be tightly regulated to ensure an immunological balance. As microRNAs (miRNAs) are critical regulators of signaling pathways, we aimed to identify specific miRNAs implicated in controlling Th17 differentiation. We were able to create a regulatory network model of murine Th cell differentiation by combining Affymetrix mRNA and miRNA arrays and in-silico analysis. In this model, the miR-212~132 and miR-182~183 clusters were significantly up-regulated upon Th17 differentiation, while the entire miR-106~363 cluster was down regulated and predicted to target well-known Th17 cell differentiation pathways. In-vitro transfection of miR-18b, miR-106 and miR-363 into primary murine CD4(+) lymphocytes decreased expression of retinoid-related orphan receptor c (Rorc), Rora, Il17a and Il17f, and abolished secretion of Th17 mediated Il17-a. Moreover, we demonstrated target site-specific regulation of the Th17 transcription factors Rora and nuclear factor of activated T-cells (Nfat) 5 by miR-18b, miR-106a and miR-363-3p through luciferase reporter assays. Here, we provide evidence that miRNAs are involved in controlling the differentiation and function of T-helper cells, offering useful tools to study and modify Th17 mediated inflammation.
AU  - Kaestle, M.
AU  - Bartel, S.
AU  - Geillinger-Kästle, K.*
AU  - Irmler, M.
AU  - Beckers, J.
AU  - Ryffel, B.*
AU  - Eickelberg, O.
AU  - Krauss-Etschmann, S.
C1  - 51353
C2  - 43181
CY  - Hoboken
SP  - 402-413
TI  - microRNA cluster 106a~363 is involved in T-Helper 17 (Th17) cell differentiation.
JO  - Immunology
VL  - 152
IS  - 3
PB  - Wiley
PY  - 2017
SN  - 0019-2805
ER  - 

TY  - JOUR
AU  - Karo-Atar, D.*
AU  - Moskovitz, I.*
AU  - Eickelberg, O.
AU  - Königshoff, M.
AU  - Munitz, A.*
C1  - 47035
C2  - 40480
SP  - 278-279
TI  - Paired immunoglobulin-like receptor B (PIR-B) negatively regulates pulmonary fibrosis by suppressing IL-4-induced macrophage activation.
JO  - Immunology
VL  - 137
PY  - 2012
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - Adoptive transfer of T cells genetically modified with tumour-specific T-cell receptors (TCR) is a promising novel approach in the treatment of cancer. We have previously isolated an allorestricted MHC class I-restricted TCR with specificity for Formin-like protein 1 (FMNL1) with potent activity against chronic lymphocytic leukaemia cells. CD4(+) T cells have been described to be highly important for tumour elimination although TCR derived from CD4(+) T cells with anti-tumour reactivity have been only rarely described. In this study we aimed to isolate MHC class-II-restricted CD4(+) T cells and TCR with specificity for leukaemia antigens. We used professional antigen-presenting cells pulsed with the leukaemia-associated and tumour-associated antigen FMNL1 for stimulation of autologous T cells in vitro. We isolated two CD4(+) HLA-DR-restricted T-cell clones and T-cell-derived TCR with so far unknown specificity but high reactivity against lymphoma cells and native malignant cells derived from HLA-matched patients with diverse leukaemias. Moreover, characterization of the TCR after TCR gene transfer revealed that specific characteristics of isolated TCR as reactivity in response to Toll-like receptors were transferable on effector cells. Our results have a major impact on the development of novel immunotherapies. They demonstrate that TCR with potent HLA-DR-restricted anti-leukaemic reactivity against so far undefined self-restricted antigens can be isolated from the healthy autorestricted CD4(+) T-cell repertoire and these TCR are highly interesting candidate tools for novel immunotherapies.
AU  - Weigand, L.U.*
AU  - Liang, X.*
AU  - Schmied, S.*
AU  - Mall, S.*
AU  - Klar, R.*
AU  - Stötzer, O.J.*
AU  - Salat, C.*
AU  - Götze, K.*
AU  - Mautner, J.
AU  - Peschel, C.*
AU  - Krackhardt, A.M.
C1  - 10869
C2  - 30374
SP  - 226-238
TI  - Isolation of human MHC class II-restricted T cell receptors from the autologous T-cell repertoire with potent anti-leukaemic reactivity.
JO  - Immunology
VL  - 137
IS  - 3
PB  - WILEY-BLACKWELL
PY  - 2012
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - Chronic hepatitis B virus (HBV) infection is the result of an inadequate immune response towards the virus. Myeloid dendritic cells (mDC) of patients with chronic HBV are impaired in their maturation and function, resulting in more tolerogenic rather than immunogenic responses, which may contribute to viral persistence. The mechanism responsible for altered mDC function remains unclear. The HBV-infected patients display large amounts of HBV particles and viral proteins in their circulation, especially the surface antigen HBsAg, which allows multiple interactions between the virus, its viral proteins and DC. To assess whether HBV directly influences mDC function, the effects of HBV and HBsAg on human mDC maturation and function were investigated in vitro. As already described for internalization of HBV by DC, the present study shows that peripheral blood-derived mDC of healthy controls also actively take up HBsAg in a time-dependent manner. Cytokine-induced maturation in the presence of HBV or HBsAg resulted in a significantly more tolerogenic mDC phenotype as demonstrated by a diminished up-regulation of costimulatory molecules and a decreased T-cell stimulatory capacity, as assessed by T-cell proliferation and interferon-gamma production. In addition, the presence of HBV significantly reduced interleukin-12 production by mDC. These results show that both HBV particles and purified HBsAg have an immune modulatory capacity and may directly contribute to the dysfunction of mDC in patients with chronic HBV. The direct immune regulatory effect of HBV and circulating HBsAg particles on the function of DC can be considered as part of the mechanism by which HBV escapes immunity.
AU  - Op den Brouw, M.L.*
AU  - Binda, R.S.*
AU  - van Roosmalen, M.H.*
AU  - Protzer, U.
AU  - Janssen, H.L.*
AU  - van der Molen, R.G.*
AU  - Woltman, A.M.*
C1  - 3495
C2  - 25793
SP  - 280-289
TI  - Hepatitis B virus surface antigen impairs myeloid dendritic cell function: A possible immune escape mechanism of hepatitis B virus.
JO  - Immunology
VL  - 126
IS  - 2
PB  - Blackwell
PY  - 2009
SN  - 0019-2805
ER  - 

TY  - JOUR
AU  - Köllisch, G.
AU  - Kalali, N.B.*
AU  - Voelcker, V.*
AU  - Wallich, R.*
AU  - Behrendt, H.
AU  - Ring, J.*
AU  - Bauer, S.*
AU  - Jakob, T.
AU  - Mempel, M.
AU  - Ollert, M.*
C1  - 5403
C2  - 22966
SP  - 531-541
TI  - Various members of the Toll-like receptor family contribute to the innate immune response of human epidermal keratinocytes.
JO  - Immunology
VL  - 114
PY  - 2005
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - Gene targeting at the immunoglobulin loci of B cells is an efficient tool for studying immunoglobulin expression or generating chimeric antibodies. We have shown that vector integration induced by human immunoglobulin G1 (IgG1) insertion vectors results in subsequent vector excision mediated by the duplicated target sequence, whereas replacement events which could be induced by the same constructs remain stable. We could demonstrate that the distribution of the vector homology strongly influences the genetic stability obtained. To this end we developed a novel type of a heavy chain replacement vector making use of the heavy chain class switch recombination sequence. Despite the presence of a two-sided homology this construct is universally applicable irrespective of the constant gene region utilized by the B cell. In comparison to an integration vector the frequency of stable incorporation was strongly increased, but we still observed vector excision, although at a markedly reduced rate. The latter events even occurred with circular constructs. Linearization of the construct at various sites and the comparison with an integration vector that carries the identical homology sequence, but differs in the distribution of homology, revealed the following features of homologous recombination of immunoglobulin genes: (i) the integration frequency is only determined by the length of the homology flank where the cross-over takes place; (ii) a 5' flank that does not meet the minimum requirement of homology length cannot be complemented by a sufficient 3' flank; (iii) free vector ends play a role for integration as well as for replacement targeting; (iv) truncating recombination events are suppressed in the presence of two flanks. Furthermore, we show that the switch region that was used as 3' flank is non-functional in an inverted orientation.
AU  - Kardinal, C.
AU  - Selmayr, M.
AU  - Mocikat, R.
C1  - 58139
C2  - 0
SP  - 309-315
TI  - Genetic stability of gene targeted immunoglobulin loci. I. Heavy chain isotype exchange induced by a universal gene replacement vector.
JO  - Immunology
VL  - 89
IS  - 3
PY  - 1996
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - Chimeric antibodies against the murine T-cell antigen Thy-1.2 were generated in amounts sufficient for in vivo studies by substituting the constant gene segments via homologous recombination in the hybridoma cell. We show that an integration vector targets the heavy chain locus at high frequency even in a non-isogenic situation. Using this vector type, for the first time expression rates were obtained that were identical to the parental hybridoma. The use of the gpt selection marker seems to be crucial for efficient expression, and may overcome a recently claimed drawback of vector integration. A chimeric antibody produced by gene targeting was characterized in vitro and in vivo.
AU  - Mocikat, R.
AU  - Kardinal, C.
AU  - Lang, P.
AU  - Zeidler, R.
AU  - Thierfelder, S.
C1  - 27610
C2  - 32765
SP  - 159-163
TI  - Unaltered immunoglobulin expression in hybridoma cells modified by targeting of the heavy chain locus with an integration vector.
JO  - Immunology
VL  - 84
IS  - 1
PY  - 1995
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - This study deals with the question of whether mouse bone marrow-derived mast cells are able to produce interleukin-6 (IL-6) in vitro. For this purpose, a panel of primary mast cell clones from limiting-dilution microcultures, of permanent IL-3-dependent mast cell lines and autonomous malignant sublines, was screened. All of these lines were found to produce growth factor activity for IL-6-dependent mouse hybridoma cells (7TD1), which could be completely neutralized by the monoclonal anti-IL-6-antibody 6B4. Transcriptional activity of the IL-6 gene was demonstrated in both IL-3-dependent mast cells and autonomous sublines using a mouse IL-6-specific cDNA probe.
AU  - Hültner, L.
AU  - Szöts, H.
AU  - Welle, M.
AU  - van Snick, J.
AU  - Moeller, J.
AU  - Dörmer, P.
C1  - 17823
C2  - 10768
SP  - 408-413
TI  - Mouse Bone Marrow-Derived IL-3-dependent Mast Cells and Autonomous Sublines Produce IL-6.
JO  - Immunology
VL  - 67
IS  - 3
PY  - 1989
SN  - 0019-2805
ER  - 

TY  - JOUR
AB  - This study deals with the question of whether mouse bone marrow-derived mast cells are able to produce interleukin-6 (IL-6) in vitro. For this purpose, a panel of primary mast cell clones from limiting-dilution microcultures, of permanent IL-3-dependent mast cell lines and autonomous malignant sublines, was screened. All of these lines were found to produce growth factor activity for IL-6-dependent mouse hybridoma cells (7TD1), which could be completely neutralized by the monoclonal anti-IL-6-antibody 6B4. Transcriptional activity of the IL-6 gene was demonstrated in both IL-3-dependent mast cells and autonomous sublines using a mouse IL-6-specific cDNA probe.
AU  - Hültner, L.
AU  - Szöts, H.
AU  - Welle, M.M.
AU  - Van Snick, J.L.
AU  - Moeller, J.
AU  - Dörmér, P.G.
C1  - 41702
C2  - 0
SP  - 408-413
TI  - Mouse bone marrow-derived IL-3 dependent mast cells and autonomous sublines produce IL-6.
JO  - Immunology
VL  - 67
IS  - 3
PY  - 1989
SN  - 0019-2805
ER  -