TY  - JOUR
AB  - Dendritic cells (DC) and T-cells are mediators of CTL-responses. Autologous (from patients with acute myeloid leukaemia (AML) or myelodysplasia (MDS)) or allogeneic (donor)-T-cells stimulated by DCleu, gain an efficient lysis of naive blasts, although not in every case. CXCL8, -9, -10, CCL2, -5 and Interleukin (IL-12) were quantified by Cytometric Bead Array (CBA) in supernatants from 5 DC-generating methods and correlated with AML-/MDS-patients' serum-values, DC-/T-cell-interactions/antileukemic T-cell-reactions after mixed lymphocyte culture (MLC) and patients' clinical course. The blast-lytic activity of T-cells stimulated with DC or mononuclear cells (MNC) was quantified in a cytotoxicity assay. Despite great variations of chemokine-levels, correlations with post-stimulation (after stimulating T-cells with DC in MLC) improved antileukemic T-cell activity were seen: higher released chemokine-values correlated with improved T-cells' antileukemic activity (compared to stimulation with blast-containing MNC) - whereas with respect to the corresponding serum values higher CXCL8-, -9-, and -10- but lower CCL5- and -2-release correlated with improved antileukemic activity of DC-stimulated (vs. blast-stimulated) T-cells. In DC-culture supernatants higher chemokine-values correlated with post-stimulation improved antileukemic T-cell reactivity, whereas higher serum-values of CXCL8, -9, and -10 but lower serum-values of CCL5 and -2 correlated with post-stimulation improved antileukemic T-cell-reactivity. In a context of 'DC'-stimulation (vs serum) this might point to a change of (CCL5 and -2-associated) functionality from a more 'inflammatory' or 'tumor-promoting' to a more 'antitumor'-reactive functionality. This knowledge could contribute to develop immune-modifying strategies that promote antileukemic (adaptive) immune-responses.
AU  - Merle, M.*
AU  - Fischbacher, D.*
AU  - Liepert, A.*
AU  - Grabrucker, C.*
AU  - Kroell, T.*
AU  - Kremser, A.*
AU  - Dreyssig, J.*
AU  - Freudenreich, M.*
AU  - Schuster, F.*
AU  - Borkhardt, A.*
AU  - Kraemer, D.*
AU  - Koehne, C.H.*
AU  - Kolb, H.J.
AU  - Schmid, C.*
AU  - Schmetzer, H.
C1  - 61785
C2  - 50458
CY  - Hackerbrucke 6, 80335 Munich, Germany
TI  - Conversion of AML-blasts to leukemia-derived dendritic cells (DCleu) in 'DC-culture-media' shifts correlations of released chemokines with antileukemic T-cell reactions.
JO  - Immunobiology
VL  - 226
IS  - 3
PB  - Elsevier Gmbh
PY  - 2021
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Non-classical human monocytes are characterized by high-level expression of cytokines like TNF, but the mechanisms involved are elusive. We have identified miRNAs and CpG-methylation sites that are unique to non-classical monocytes, defined via CD14 and CD16 expression levels. For down-regulated miRNAs that are linked to up-regulated mRNAs the dominant gene ontology term was intracellular signal transduction. This included down-regulated miRNA-20a-5p and miRNA-106b-5p, which both are linked to increased mRNA for the TRIM8 signaling molecule. Methylation analysis revealed 16 hypo-methylated CpG sites upstream of 14 differentially increased mRNAs including 2 sites upstream of TRIM8. Consistent with a positive role in signal transduction, high TRIM8 levels went along with high basal TNF mRNA levels in non-classical monocytes. Since cytokine expression levels in monocytes strongly increase after stimulation with toll-like-receptor ligands, we have analyzed non-classical monocytes (defined via slan expression) after stimulation with lipopolysaccharide (LPS). LPS-stimulated cells continued to have low miRNA-20a and miRNA-106b and high TRIM8 mRNA levels and they showed a 10-fold increase in TNF mRNA. These data suggest that decreased miRNAs and CpG hypo-methylation is linked to enhanced expression of TRIM8 and that this can contribute to the increased TNF levels in non-classical human monocytes.
AU  - Zhang, L.*
AU  - Hofer, T.P.
AU  - Zawada, A.M.*
AU  - Rotter, B.*
AU  - Krezdorn, N.*
AU  - Nößner, E.
AU  - Devaux, Y.*
AU  - Heine, G.*
AU  - Ziegler-Heitbrock, L.*
C1  - 59188
C2  - 48621
CY  - Hackerbrucke 6, 80335 Munich, Germany
TI  - Epigenetics in non-classical monocytes support their pro-inflammatory gene expression.
JO  - Immunobiology
VL  - 225
IS  - 3
PB  - Elsevier Gmbh
PY  - 2020
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Among the three human monocyte subsets, intermediate CD14++CD16+ monocytes have been characterized as particularly proinflammatory cells in experimental studies and as potential biomarkers of cardiovascular risk in clinical cohorts. To further substantiate the distinct role of intermediate monocytes within human monocyte heterogeneity, we assessed subset-specific expression of miRNAs as central epigenetic regulators of gene expression. We hypothesized that intermediate monocytes have a distinct miRNA profile compared to classical and non-classical monocytes.By using small RNA-seq we analyzed 662 miRNAs in the three monocyte subsets. We identified 38 miRNAs that are differentially expressed in intermediate monocytes compared to both classical and non-classical monocytes with a p value of <10-10, of which two miRNAs - miR-6087 (upregulated) and miR-150-5p (downregulated) - differed in their expression more than ten-fold. Pathway analysis of the 38 differentially expressed miRNAs linked intermediate monocytes to distinct biological processes such as gene regulation, cell differentiation, toll-like receptor signaling as well as antigen processing and presentation. Moreover, differentially expressed miRNAs were connected to those genes that we previously identified as markers of intermediate monocytes.In aggregation, we provide first genome-wide miRNA data in the context of monocyte heterogeneity, which substantiate the concept of monocyte trichotomy in human immunity. The identification of miRNAs that are specific for intermediate monocytes may allow to develop strategies, which particularly target this cell population while sparing the other two subsets.
AU  - Zawada, A.M.*
AU  - Zhang, L.*
AU  - Emrich, I.E.*
AU  - Rogacev, K.S.*
AU  - Krezdorn, N.*
AU  - Rotter, B.*
AU  - Fliser, D.*
AU  - Devaux, Y.*
AU  - Ziegler-Heitbrock, L.
AU  - Heine, G.H.*
C1  - 50238
C2  - 42236
CY  - Jena
SP  - 587-596
TI  - MicroRNA profiling of human intermediate monocytes.
JO  - Immunobiology
VL  - 222
IS  - 3
PB  - Elsevier Gmbh, Urban & Fischer Verlag
PY  - 2017
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Crown Copyright © 2016. Published by Elsevier GmbH. All rights reserved. Among the three human monocyte subsets, intermediate CD14++CD16+ monocytes have been characterized as particularly proinflammatory cells in experimental studies and as potential biomarkers of cardiovascular risk in clinical cohorts. To further substantiate the distinct role of intermediate monocytes within human monocyte heterogeneity, we assessed subset-specific expression of miRNAs as central epigenetic regulators of gene expression. We hypothesized that intermediate monocytes have a distinct miRNA profile compared to classical and non-classical monocytes. By using small RNA-seq we analyzed 662 miRNAs in the three monocyte subsets. We identified 38 miRNAs that are differentially expressed in intermediate monocytes compared to both classical and non-classical monocytes with a p value of < 10-10, of which two miRNAs - miR-6087 (upregulated) and miR-150-5p (downregulated) - differed in their expression more than ten-fold. Pathway analysis of the 38 differentially expressed miRNAs linked intermediate monocytes to distinct biological processes such as gene regulation, cell differentiation, toll-like receptor signaling as well as antigen processing and presentation. Moreover, differentially expressed miRNAs were connected to those genes that we previously identified as markers of intermediate monocytes. In aggregation, we provide first genome-wide miRNA data in the context of monocyte heterogeneity, which substantiate the concept of monocyte trichotomy in human immunity. The identification of miRNAs that are specific for intermediate monocytes may allow to develop strategies, which particularly target this cell population while sparing the other two subsets.
AU  - Zawada, A.M.*
AU  - Zhang, L.*
AU  - Emrich, I.E.*
AU  - Rogacev, K.S.*
AU  - Krezdorn, N.*
AU  - Rotter, B.*
AU  - Fliser, D.*
AU  - Devaux, Y.*
AU  - Ziegler-Heitbrock, L.
AU  - Heine, G.H.*
C1  - 51300
C2  - 43038
CY  - Jena
SP  - 831-840
TI  - Reprint of: MicroRNA profiling of human intermediate monocytes.
JO  - Immunobiology
VL  - 222
IS  - 6
PB  - Elsevier Gmbh, Urban & Fischer Verlag
PY  - 2017
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Tumor-derived microvesicles (TMV) can mimic effects of tumor cells leading to an increased anti-inflammatory cytokine production, such as interleukin 10 (IL-10), by tumor-infiltrating monocytes and macrophages. Yet, the mechanism of IL-10 induction by TMV in monocytes remains unclear. The co-incubation of TMV derived from the human pancreas carcinoma cell line (HPC-4) with human monocytes resulted in a nearly 30-fold increase in IL-10 protein production. This effect operates at the level of transcription since monocytes transduced with an adenovirus containing IL-10-promoter luciferase reporter gene showed a 5-fold induction of luciferase activity after treatment with TMV. Since tumor cells can express hyaluronan (HA), which participates in tumor invasion and metastases, we have tested its effect on IL-10 expression. We showed that HA at the concentration of 100μg/ml induces IL-10 protein expression and the IL-10 promoter activation in monocytes. Moreover, hyaluronidase treatment of TMV reduced IL-10 protein production by 50% and promoter activity by 40%. Inhibitors of the PI3K/Akt/mTOR pathway reduced both, TMV-induced IL-10 promoter activity and protein production, and the same was observed in monocytes when stimulated by HPC-4 cells or HA. Inhibition of PI3K activity down-regulated phosphorylation of the Akt and (to a lesser extent) mTOR proteins in monocytes following TMV or HA stimulation. When comparing monocyte subsets, TMV induced IL-10 protein and mRNA synthesis only in classical CD14(++)CD16(-) but not in CD16-positive monocytes. Our data show that TMV induce IL-10 synthesis in human classical monocytes via HA, which, in turn, activates the PI3K/Akt/mTOR pathway.
AU  - Lenart, M.*
AU  - Rutkowska-Zapala, M.*
AU  - Baj-Krzyworzeka, M.*
AU  - Szatanek, R.*
AU  - Węglarczyk, K.*
AU  - Smallie, T.*
AU  - Ziegler-Heitbrock, L.
AU  - Zembala, M.*
AU  - Siedlar, M.*
C1  - 46424
C2  - 37542
CY  - Jena
SP  - 1-10
TI  - Hyaluronan carried by tumor-derived microvesicles induces IL-10 production in classical CD14++ CD16- monocytes via PI3K/Akt/mTOR-dependent signalling pathway.
JO  - Immunobiology
VL  - 222
IS  - 1
PB  - Elsevier Gmbh, Urban & Fischer Verlag
PY  - 2015
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - T-cells play an important role in the remission-maintenance in AML-patients (pts) after SCT, however the role of LAA- (WT1, PR1, PRAME) or minor-histocompatibility (mHag, HA1) antigen-specific CD4(+) and CD8(+)T-cells is not defined. A LAA/HA1-peptide/protein stimulation, cloning and monitoring strategy for specific CD8(+)/CD4(+)T-cells in AML-pts after SCT is given. Our results show that (1) LAA-peptide-specific CD8+T-cells are detectable in every AML-pt after SCT. CD8(+)T-cells, recognizing two different antigens detectable in 5 of 7 cases correlate with long-lasting remissions. Clonal TCR-Vβ-restriction exemplarily proven by spectratyping in PRAME-specific CD8(+)T-cells; high PRAME-peptide-reactivity was CD4(+)-associated, as shown by IFN-γ-release. (2) Two types of antigen-presenting cells (APCs) were tested for presentation of LAA/HA1-proteins to CD4(+)T-cells: miniEBV-transduced lymphoblastoid cells (B-cell-source) and CD4-depleted MNC (source for B-cell/monocyte/DC). We provide a refined cloning-system for proliferating, CD40L(+)CD4(+)T-cells after LAA/HA1-stimulation. CD4(+)T-cells produced cytokines (GM-CSF, IFN-γ) upon exposure to LAA/HA1-stimulation until after at least 7 restimulations and demonstrated cytotoxic activity against naive blasts, but not fibroblasts. Antileukemic activity of unstimulated, stimulated or cloned CD4(+)T-cells correlated with defined T-cell-subtypes and the clinical course of the disease. In conclusion we provide immunological tools to enrich and monitor LAA/HA1-CD4(+)- and CD8(+)T-cells in AML-pts after SCT and generate data with relevant prognostic value. We were able to demonstrate the presence of LAA-peptide-specific CD8(+)T-cell clones in AML-pts after SCT. In addition, we were also able to enrich specific antileukemic reactive CD4(+)T-cells without GvH-reactivity upon repeated LAA/HA1-protein stimulation and limiting dilution cloning.
AU  - Steger, B.
AU  - Milosevic, S.
AU  - Doessinger, G.*
AU  - Reuther, S.*
AU  - Liepert, A.*
AU  - Braeu, M.
AU  - Schick, J.*
AU  - Vogt, V.*
AU  - Schuster, F.*
AU  - Kroell, T.*
AU  - Busch, D.H.
AU  - Borkhardt, A.*
AU  - Kolb, H.-J.
AU  - Tischer, J.*
AU  - Buhmann, R.
AU  - Schmetzer, H.
C1  - 28638
C2  - 33510
CY  - Jena
SP  - 247-260
TI  - CD4+and CD8+T-cell reactions against leukemia-associated- or minor-histocompatibility-antigens in AML-patients after allogeneic SCT.
JO  - Immunobiology
VL  - 219
IS  - 4
PB  - Elsevier
PY  - 2014
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - The CD16-positive monocytes have been first described in 1988 but to date no selective defect in the number of these cells in blood has been reported. We now describe a family in which three of four siblings lack both CD16-positive monocyte subsets, i.e. the nonclassical and the intermediate monocytes. All three had CD16-positive monocytes of 2cells/μl or less as compared to 52±18cells/μl in healthy controls. The index case was affected by recurrent pleural effusion and infections and had evidence of an auto-inflammatory condition but no mutation of any of the relevant candidate genes. The other two siblings without CD16-positive monocytes were apparently healthy. There was no defect in serum M-CSF levels and no mutation in the M-CSF and M-CSFR genes. The data indicate that the absence of CD16-positive monocytes in blood does not lead to disease.
AU  - Frankenberger, M.
AU  - Ekici, A.B.*
AU  - Angstwurm, M.W.*
AU  - Hoffmann, H.*
AU  - Hofer, T.P.
AU  - Heimbeck, I.
AU  - Meyer, P.
AU  - Lohse, P.*
AU  - Wjst, M.
AU  - Häussinger, K.*
AU  - Reis, A.*
AU  - Ziegler-Heitbrock, L.
C1  - 7393
C2  - 29700
SP  - 169-174
TI  - A defect of CD16-positive monocytes can occur without disease.
JO  - Immunobiology
VL  - 218
IS  - 2
PB  - Elsevier
PY  - 2013
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - The success of a vaccine consists in the induction of an innate immune response and subsequent activation of the adaptive immune system. Because antigens are usually not immunogenic, the addition of adjuvants that activate innate immunity is required. The mycobacterial cord factor trehalose-6,6'-dimycolate (TDM) and its synthetic adjuvant analogue trehalose-6,6'-dibehenate (TDB) rely on the C-type lectin Mincle and the signaling molecules Syk and Card9 to trigger innate immunity. In this study, we show that stimulation of bone marrow-derived dendritic cells (BMDCs) with TDB induces Nlrp3 inflammasome-dependent IL-1β secretion. While Card9 is required for NF-κB activation by TDB, it is dispensable for TDB-induced activation of the Nlrp3 inflammasome. Additionally, efflux of intracellular potassium, lysosomal rupture, and oxygen radical (ROS) production are crucial for caspase-1 processing and IL-1β secretion by TDB. In an in vivo inflammation model, we demonstrate that the recruitment of neutrophils by TDB is significantly reduced in the Nlrp3-deficient mice compared to the wild-type mice, while the production of chemokines in vitro is not influenced by the absence of Nlrp3. These results identify the Nlrp3 inflammasome as an essential mediator for the induction of an innate immune response triggered by TDB.
AU  - Schweneker, K.*
AU  - Gorka, O.*
AU  - Schweneker, M.*
AU  - Poeck, H.*
AU  - Tschopp, J.*
AU  - Peschel, C.*
AU  - Ruland, J.
AU  - Groß, O.*
C1  - 11170
C2  - 30532
SP  - 664-673
TI  - The mycobacterial cord factor adjuvant analogue trehalose-6,6'-dibehenate (TDB) activates the Nlrp3 inflammasome.
JO  - Immunobiology
VL  - 218
IS  - 4
PB  - Elsevier
PY  - 2013
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Background: Donor lymphocyte transfusion (DLT) may induce the graft-versus-leukemia (GVL) effect for patients with AML relapsed after transplant. However, AML is a highly diverse disease and the limited overall efficacy of DLT in clinical practice emphasizes the importance of identifying a specific subgroup of patients who might benefit from this treatment approach. Objective: To monitor the cellular immune response after DLT, we developed an active specific immunization strategy using in vitro generated AML-trained T cells to induce a highly specific antileukemic T-cell response and thus established a novel nonradioactive assay system to assess the antileukemia immunity by flow cytometry, correlated with [3H]-thymidine uptake. Methods: The myeloid blasts derived from five patients with AML relapsed post-allogeneic hematopoietic stem cell transplantation (allo-HSCT) were first labeled with CFDA (5,6-carboxyfluorescein diacetate succinimidyl ester). To analyze the growth inhibitory potential of the donor T cells trained by AML progenitor cells, the myeloid blasts were induced to proliferate by means of a cytokine cocktail (50 ng/mL of SCF; 25 ng/mL of IL-3; 100 ng/mL of GM-CSF; 100 ng/mL of G-CSF; 2 U/mL of EPO; 0.47 g/L of transferrin; and 5 x 10(-5) mmol/L of 2-ME). The T cell mediated growth inhibitory potential was detected after 5 days by flow cytometry and correlated with [3H]-thymidine uptake. The simultaneous use of TO-PRO-dye and calibrate beads allowed not only the cell viability to be known but also allowed quantification of the effector function. Results: Here, we applied a CFDA dye to track the proliferation and expansion of AML blasts in response to the cytokine cocktail in vitro. AML-trained T cells, expressed high levels of the activation markers CD25 and CD69, and were generated to recognize the leukemic progenitor cells and inhibit cytokine-induced leukemic cell proliferation, which is an active specific immunization strategy circumventing the identification of leukemia-associated antigens. The capability of proliferation inhibition of AML-trained T cells evaluated with our nonradioactive, CFDA-based assay provided comparable results with the classic [3H]]-thymidine assay with an even lower ratio of effector to target cells. Conclusion: Taken together, the novel, nonradioactive, CFDA-based assay was a robust tool to monitor the antileukemic immune response after DLT in myeloid leukemias.
AU  - Yang, T.
AU  - Chen, Z.-Z.*
AU  - Kolb, H.-J.
AU  - Buhmann, R.
C1  - 24874
C2  - 31740
SP  - 548-553
TI  - A novel nonradioactive CFDA assay to monitor the cellular immune response in myeloid leukemia.
JO  - Immunobiology
VL  - 218
IS  - 4
PB  - Elsevier
PY  - 2013
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - All-trans retinoic acid (ATRA) is a potent retinoid, which has been used successfully in different clinical settings as a potential drug to treat COPD and emphysema. In the present study, we analyzed genes modulated by ATRA by performing mRNA expression array analysis on alveolar macrophages after treatment with ATRA. Here we observed a 375-fold up-regulation of Prostaglandin-E Synthase (microsomal PGES-1, NM_004878 PTGES) which mediates the conversion of prostaglandin H(2) (PGH(2)) to Prostaglandin E(2) (PGE(2)). We furthermore studied the expression of PTGES after treatment with ATRA in human monocyte-derived macrophages (MDMs) and bronchoalveolar lavage (BAL) cells. ATRA up-regulated PTGES mRNA expression in MDMs generated with M-CSF by 2500-fold whereas in M-CSF+IL-13 macrophages the up-regulation was only 20-fold. Similarly, ATRA up-regulated PTGES mRNA expression by factor 1524 in BAL cells. The up-regulation of PTGES mRNA expression by ATRA is both time and dose dependent. IL-13 suppressed the ATRA induced PTGES expression at both mRNA and protein level in MDM and BAL cells. We also observed that LPS acts synergistically with ATRA in MDMs and strongly induces PTGES expression. ATRA had little impact on cyclooxygenase-1 and -2 (COX-1 and -2) expression as compared to PTGES expression under the same experimental conditions. Furthermore, we observed an induction of PGE(2) levels by ATRA in BAL cells. These data indicate that ATRA is a potent inducer of PTGES expression in human macrophages but not in alternatively activated macrophages and suggest that the eicosanoid pathway is important for ATRA action in macrophages.
AU  - Mamidi, S.
AU  - Hofer, T.P.
AU  - Hoffmann, R.*
AU  - Ziegler-Heitbrock, L.
AU  - Frankenberger, M.
C1  - 7413
C2  - 29709
SP  - 593-600
TI  - All-trans retinoic acid up-regulates Prostaglandin-E Synthase expression in human macrophages.
JO  - Immunobiology
VL  - 217
IS  - 6
PB  - Elsevier
PY  - 2012
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - The endocannabinoid system (ECS) consists of two cannabinoid (CB) receptors, namely CB(1) and CB(2) receptor, and their endogenous (endocannabinoids) and exogenous (cannabinoids, e.g. delta-9-tetrahydrocannabinol (THC)) ligands which bind to these receptors. Based on studies suggesting a role of THC and the ECS in inflammation, the objective of this study was to examine their involvement in type I hypersensitivity using a murine model of allergic airway inflammation. THC treatment of C57BL/6 wildtype mice dramatically reduced airway inflammation as determined by significantly reduced total cell counts in bronchoalveolar lavage (BAL). These effects were greatest when mice were treated during both, the sensitization and the challenge phase. Furthermore, systemic immune responses were significantly suppressed in mice which received THC during sensitization phase. To investigate a role of CB(1/2) receptors in this setting, we used pharmacological blockade of CB(1) and/or CB(2) receptors by the selective antagonists and moreover CB(1)/CB(2) receptor double-knockout mice (CB(1)(-/-)/CB(2)(-/-)) and found neither significant changes in the cell patterns in BAL nor in immunoglobulin levels as compared to wildtype mice. Our results indicate that the activation of the ECS by applying the agonist THC is involved in the development of type I allergies. However, CB(1)/CB(2) receptor-independent signalling seems likely in the observed results.
AU  - Braun, A.
AU  - Engel, T.
AU  - Aguilar-Pimentel, J.A.
AU  - Zimmer, A.*
AU  - Jakob, T.*
AU  - Behrendt, H.
AU  - Mempel, M.
C1  - 5266
C2  - 27878
SP  - 466-476
TI  - Beneficial effects of cannabinoids (CB) in a murine model of allergen-induced airway inflammation: Role of CB(1)/CB(2) receptors.
JO  - Immunobiology
VL  - 216
IS  - 4
PB  - Elsevier
PY  - 2011
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - This study tested the hypothesis that prolonged severe hypoxia during monocyte to macrophage differentiation results in macrophages with a pattern of gene expression and phenotype distinct from those maturing in normal oxygen levels. Macrophages accumulate in hypoxic and anoxic areas within pathological sites such as tumours, wounds, and arthritic joints, and have been proposed as vehicles for gene therapy delivery to such tissues. Several non-pathological tissues are also hypoxic. We therefore argue that differentiation from monocyte to macrophage in hypoxic conditions is a common occurrence. However, the effect of long term severe hypoxia on monocyte to macrophage differentiation has not been studied. Here, using primary human peripheral blood monocytes, we show that maturation for 5 days in 0.2% oxygen results in decreased phagocytosis, and decreased CD40 and CD206 expression. Chronic hypoxia induced much higher mRNA levels of the pro-angiogenic cytokine, VEGF, in adherence-purified macrophages (27-fold), CD14-magnetic bead purified monocytes (90-fold), and PBMC (104-fold) compared to acute (24h) hypoxia (11, 17 and 9-fold, respectively). This suggests that macrophages may play an even greater role in angiogenesis than previously appreciated. Furthermore, chronic hypoxia resulted in up-regulation of HIF-1α mRNA, in all monocyte-derived macrophage types studied. Actinomycin D experiments indicate that the increases in HIF-1α mRNA were not due to increased mRNA stability. To our knowledge this is the first study demonstrating up-regulation of HIF-1α mRNA by hypoxia in macrophages. Taken together, the data support the hypothesis that hypoxia affects monocyte to macrophage maturation, resulting in a distinct gene expression pattern and phenotype.
AU  - Staples, K.J.*
AU  - Sotoodehnejadnematalahi, F.*
AU  - Pearson, H.*
AU  - Frankenberger, M.
AU  - Francescut, L.*
AU  - Ziegler-Heitbrock, L.
AU  - Burke, B.*
C1  - 5142
C2  - 28566
SP  - 832-839
TI  - Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA.
JO  - Immunobiology
VL  - 216
IS  - 7
PB  - Elsevier
PY  - 2011
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Equine recurrent uveitis serves as a spontaneous model for human autoimmune uveitis. Unpredictable relapses and ongoing inflammation in the eyes of diseased horses as well as in humans lead to destruction of the retina and finally result in blindness. However, the molecular mechanisms leading to inflammation and retinal degeneration are not well understood. An initial screening for differentially regulated proteins in sera of uveitic cases compared to healthy controls revealed an increase of the alternative pathway complement component factor B in ERU cases. To determine the activation status of the complement system, sera were subsequently examined for complement split products. We could demonstrate a significant higher concentration of the activation products B/Ba, B/Bb, Bb neoantigen, iC3b and C3d in uveitic condition compared to healthy controls, whereas for C5b-9 no differences were detected. Additionally, we investigated complement activation directly in the retina by immunohistochemistry, since it is the main target organ of this autoimmune disease. Interestingly, infiltrating cells co-expressed activated factor Bb neoantigen, complement split product C3d as well as CD68, a macrophage marker. In this study, we could demonstrate activation of the complement system both systemically as well as in the eye, the target organ of spontaneous recurrent uveitis. Based on these novel findings, we postulate a novel role for macrophages in connection with complement synthesis at the site of inflammation.
AU  - Zipplies, J.K.*
AU  - Kirschfink, M.*
AU  - Amann, B.*
AU  - Hauck, S.M.
AU  - Stangassinger, M.*
AU  - Deeg, C.A.*
C1  - 4973
C2  - 27749
SP  - 949-955
TI  - Complement factor B expression profile in a spontaneous uveitis model.
JO  - Immunobiology
VL  - 215
IS  - 12
PB  - Elsevier
PY  - 2010
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Bronchiolitis obliterans with organizing pneumonia (BOOP) is a disease affecting small airways and alveoli. It is characterized by interstitial inflammation rich in foamy macrophages and by fibroblastic connective tissue expanding into the airway and alveolar lumen. We report herein on a 54-year-old male BOOP patient who was treated with glucocorticoids (GCs) and who over a 5-year period had three relapses. At diagnosis the patient showed elevated CD14(+)CD16(+) monocyte numbers (85cells/mul) and increased serum C-reactive protein (CRP) levels (29.4mg/l). With GC therapy both parameters decreased within a few days. Diagnosis of relapse was preceded by a rise in CD14(+)CD16(+) monocyte numbers and in CRP levels which again responded to GC treatment. We conclude that determination of CD14(+)CD16(+) monocytes is a useful marker for monitoring of BOOP diagnosis and GC therapy.
AU  - Fertl, A.*
AU  - Menzel, M.*
AU  - Hofer, T.P.
AU  - Morresi-Hauf, A.*
AU  - Ziegler-Heitbrock, L.
AU  - Frankenberger, M.
C1  - 718
C2  - 25672
SP  - 909-916
TI  - Monitoring of glucocorticoid therapy by assessment of CD14(+)CD16(+) monocytes: A case report.
JO  - Immunobiology
VL  - 213
IS  - 9-10
PB  - Elsevier
PY  - 2008
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Many plant-pathogen interactions are controlled by specific interactions between pathogen avirulence (avr) gene loci and the corresponding plant resistance R locus (gene-for-gene-hypothesis). Very often, this type of interaction culminates in a hypersensitive reaction (HR). However, recently pathogen-associated molecular patterns (PAMPs) such as flagellin or lipopolysaccharides (LPS) that are common to all bacteria have been shown to act as general elicitors of basal or innate immune responses in several plant species. Here, we summarize the genetic programs in Arabidopsis thaliana behind the LPS-induced basal response and the HR induced by harpin, respectively. Using Agilent Arabidopsis cDNA microarrays consisting of approximately 15,000 oligomers, changes in transcript accumulation of treated cells were monitored over a period of 24h after elicitor treatment. Analysis of the array data revealed significant responses to LPS (309 genes), harpin (951 genes) or both (313 genes). Concentrating our analysis on the genes encoding transcription factors, defence genes, cell wall biogenesis-related genes and signal transduction components we monitored interesting parallels, but also remarkably different expression patterns. Harpin and LPS induced an overlapping set of genes involved in cell wall biogenesis, cellular communication and signalling. The pattern of induced genes associated with cell rescue and general stress responses such as small heat-shock proteins was highly similar. In contrast, there is a striking difference regarding some of the most prominent, central components of plant defence such as WRKY transcription factors and oxidative burst-associated genes like NADPH oxidases, whose expression became apparent only after treatment with harpin. While both harpin and LPS can stimulate plant immunity in Arabidopsis, the PAMP LPS induces much more subtle host reactions at the transcriptome scale. The defence machinery induced by harpin resembles the known HR-type host responses leading to cell death after treatment with this elicitor. LPS is a weak inducer of basal resistance and induces a different pattern of genes. Strikingly the biggest overlap (40) of responding genes was found between the early harpin response (30min) and the late LPS response (24h).
AU  - Livaja, M.*
AU  - Zeidler, D.*
AU  - von Rad, U.
AU  - Durner, J.
C1  - 2612
C2  - 25330
SP  - 161-171
TI  - Transcriptional responses of Arabidopsis thaliana to the bacteria-derived PAMPs harpin and lipopolysaccharide.
JO  - Immunobiology
VL  - 213
IS  - 3-4
PB  - Elsevier
PY  - 2008
SN  - 0171-2985
ER  - 

TY  - JOUR
AU  - Hofer, T.P.
AU  - Frankenberger, M.
AU  - Staples, K.J.*
AU  - Ziegler-Heitbrock, L.
C1  - 4100
C2  - 23770
SP  - 455-462
TI  - Expression of p57-Kip2 in monocytes and macrophages.
JO  - Immunobiology
VL  - 211
PY  - 2006
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Th2 cells play a central role in type I allergies. However, the source of interleukin-4 which may lead to a Th1/Th2 imbalance is unknown. Valpha24+CD161+ Natural killer T (NKT) cells secrete high amounts of interleukin-4 and/or interferon-gamma and are assumed to participate in the initiation of Th1/Th2 immune responses. Their contribution to the development of Th2-dependent type I allergies is controversial. Our objective in this paper was to determine whether Valpha24+CD161+ NKT cells differ in atopic and non-atopic adults. Venous blood was obtained from thirteen atopic and sixteen healthy adult probands. Valpha24+CD161+ NKT cells were determined in CD4+, CD8(bright/dim) and CD4-CD8- lymphocytes by flow cytometry. At the molecular level, the amounts of T cell receptor (TCR) AV24-AJ18 transcripts were quantified with respect to TCRAV24 chain transcripts alone or to all TCR alpha chain transcripts. To detect potential inserted nucleotides in the N-region, a novel real-time PCR-based technology was applied. Both CD4+ and CD4-CD8- NKT cells were present at higher frequencies than CD8+ NKT cells in all probands. CD8(dim) NKT cell levels were lower in healthy individuals, although not statistically significantly different to the patients. Amounts of AV24-AJ18 transcripts in relation to total TCR alpha-chains and to TCRAV24 alone were equal in both proband groups. N-region diversity was detected in four clones from four different individuals, but altered the amino acid sequence in only one clone of an atopic donor. Analysis of Valpha24+CD161+ NKT cell frequencies at both the cellular and molecular levels failed to reveal significant differences in peripheral blood of atopic and non-atopic probands. If NKT cells contribute to development of type I allergies they must do so at earlier times or in other locations.
AU  - Prell, C.*
AU  - Konstantopoulos, N.*
AU  - Heinzelmann, B.*
AU  - Frankenberger, B.
AU  - Reinhardt, D.*
AU  - Schendel, D.J.
AU  - Krauss-Etschmann, S.
C1  - 9524
C2  - 21566
SP  - 367-380
TI  - Frequency of Vα24+CD161+ natural killer T cells and invariant TCRAV24-AJ18 transcripts in atopic and non-atopic individuals.
JO  - Immunobiology
VL  - 208
IS  - 4
PB  - Elsevier
PY  - 2003
SN  - 0171-2985
ER  - 

TY  - JOUR
AU  - Strobl, L.J.
AU  - Höfelmayr, H.
AU  - Stein, C.
AU  - Marschall, G.
AU  - Brielmeier, M.
AU  - Laux, G.
AU  - Bornkamm, G.W.
AU  - Zimber-Strobl, U.
C1  - 9525
C2  - 22863
SP  - 299-306
TI  - Both Epstein-Barr virus nuclear antigen 2 (EBNA2) and activated notch 1 tranactivate genes ba interacting with the cellular protein RBP-Jk. 
JO  - Immunobiology
VL  - 198
PY  - 1997
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - We studied with specific polyclonal and monoclonal antibodies against human ecto-5'-nucleotidase whether the enzyme, located on the surface of human peripheral lymphocytes, could function as a mitogenic receptor for the lectins PHA, Con A and PWM. Strong, but unspecific inhibitory effects on lymphocyte stimulation are observed with unfractionated antisera and ascitic fluids. However, when purified IgG from these sources is used, no such effects are found, while at the same time, complete inhibition of ecto-5'-nucleotidase activity is maintained. It is concluded that the enzyme does not act as a mitogenic receptor for the lectins. When purine de novo synthesis of the lymphocytes is blocked by aminopterin and purine nucleotides in the extracellular medium are given as the only purine source, lymphocyte stimulation becomes dependent on the enzymatic activity of ecto-5'-nucleotidase. This is independent of the lectin used. Under these conditions, enzyme activity on the 20-30% 5'-nucleotidase-positive cells is necessary and is sufficient to support the stimulation of the whole culture. In these cultures, anti-5'-nucleotidase-IgG completely depresses cell proliferation, showing clearly that this is the only enzyme on the lymphocyte surface that is capable of degrading extracellular nucleotides.
AU  - Andree, T.H.
AU  - Gutensohn, W.
AU  - Kummer, U.
C1  - 41549
C2  - 36268
SP  - 214-225
TI  - Is ecto-5'-nucleotidase essential for stimulation of human lymphocytes? Evidence against a role of the enzyme as mitogenic lectin receptor.
JO  - Immunobiology
VL  - 175
IS  - 3
PY  - 1987
SN  - 0171-2985
ER  - 

TY  - JOUR
AB  - Human peripheral-blood mononuclear cells forming rosettes with dog and rhesus-monkey red blood cells was characterized with a double marker analysis combining immunofluorescence with a series of monoclonal antibodies and rosette formation. Dog rosette-forming cells are a minor subset of T cells which does not correspond to the helper or suppressor subsets of T cells as determined by the OKT antibodies. Dog rosette formation can be inhibited by two monoclonal antibodies directed against the receptor for sheep red blood cells on human lymphocytes. Rhesus-monkey rosettes can be formed by T lymphocytes as well as by monocytes, null cells, and B cells, as determined by the reactivities with the monoclonal antibodies OKM1 and anti-Ia. OKM1-positive cells are relatively enriched in these rosettes. No inhibition is seen with antibodies directed against the receptor for sheep red blood cells. In conclusion, we demonstrated with the use of monoclonal antibodies that dog and rhesus-monkey rosettes are qualitatively different phenomena.
AU  - Munker, R.
AU  - Stünkel, K.G.E.
C1  - 42326
C2  - 0
SP  - 78-85
TI  - Characterization with monoclonal antibodies of human lymphocytes forming dog and rhesus-monkey rosettes.
JO  - Immunobiology
VL  - 162
IS  - 1
PY  - 1982
SN  - 0171-2985
ER  -