Adaptation of Human CD4+ T Cells to Pathophysiological Hypoxia: A Transcriptome Analysis

2009 ◽  
Vol 36 (12) ◽  
pp. 2655-2669 ◽  
Author(s):  
TIMO GABER ◽  
THOMAS HÄUPL ◽  
GRIT SANDIG ◽  
KAROLINA TYKWINSKA ◽  
MONIQUE FANGRADT ◽  
...  
Keyword(s):  
T Cells ◽  
Synovial Tissue ◽  
Cd4 T Cells ◽  
Protein Modification ◽  
Quantitative Polymerase Chain Reaction ◽  
Low Oxygen ◽  
Rheumatoid Synovium ◽  
Tissue Samples ◽  
Oxygen Levels ◽  
Hypoxic Conditions

Objective.Inflamed tissues are usually characterized by low oxygen levels. We investigated whether pathophysiological hypoxia (pO2 < 1%) as found in the rheumatoid synovium modulates the transcriptome of human CD4+ T cells.Methods.We analyzed the extent to which hypoxia influences the transcriptome in the rheumatoid synovium according to a gene cluster reflecting adaptation to low oxygen levels. Hypoxia-inducible factor-1α (HIF-1α) was detected in the rheumatoid synovium using immunohistochemistry. Isolated human CD4+ T cells were exposed to hypoxia and analyzed using microarray analysis, quantitative polymerase chain reaction, and immunoblot detection.Results.In rheumatoid arthritis (RA) synovial tissue samples, hypoxia modulates the transcription profile. This profile is similar, but not identical, to that found in isolated CD4+ T cells incubated under hypoxic conditions. We show that HIF-1α is expressed in synovial tissue samples and in hypoxic CD4+ cells; and that hypoxia directly affects differential gene expression in human T cells with up to 4.8% modulation of the transcriptome. Functional genome analysis revealed substantial effects of hypoxia on immune response, transcriptional regulation, protein modification, cell growth and proliferation, and cell metabolism.Conclusion.Severe hypoxia, a feature of joint inflammation, considerably modulates the transcriptome of cells found in the rheumatoid synovium. Human CD4+ T cells adapt to hypoxic conditions mainly by HIF-1-driven effects on the transcriptome reflecting a profound influence on immune functions. Thus, hypoxia must be taken into account when therapeutically targeting inflammation.

scholarly journals SERPINB10 contributes to asthma by inhibiting the apoptosis of allergenic Th2 cells

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Yuqing Mo ◽  
Ling Ye ◽  
Hui Cai ◽  
Guiping Zhu ◽  
Jian Wang ◽  
...  
Keyword(s):  
T Cells ◽  
Allergic Asthma ◽  
Cd4 T Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Allergic Inflammation ◽  
Specific Ige ◽  
Th2 Cells ◽  
Th1 Cells ◽  
Th2 Response ◽  
Th1 And Th2

Abstract Background Serine peptidase inhibitor, clade B, member 10 (SERPINB10) contributes to allergic inflammation in asthma. However, its role in the T-helper type 2 (Th2) response of allergic asthma is not known. The goal of this study was to unveil the function of SERPINB10 in the Th2 response of allergic asthma and the mechanism by which SERPINB10 affects the viability of Th2 cells. Methods Th2 cytokines and serum levels of house dust mite (HDM)-specific IgE in bronchoalveolar lavage fluid were examined by ELISA in an HDM-induced asthma model. The number and apoptosis of Th1 and Th2 cells in mouse lungs were measured by flow cytometry. Naïve CD4 T cells from patients with asthma were cultured under appropriate polarizing conditions to generate Th1 and Th2 cells. SERPINB10 expression in polarized Th1 and Th2 cells was quantified by real-time reverse transcription-quantitative polymerase chain reaction. SERPINB10 expression was knocked down in human CD4 T cells with lentivirus. Results Knockdown of SERPINB10 expression significantly diminished HDM-induced Th2 cytokine secretion and level of HDM-specific IgE. After HDM exposure, SERPINB10-knockdown mice had diminished numbers of Th2 cells, but similar numbers of Th1 cells, compared with those in negative-control mice. Th2 cells of SERPINB10-knockdown mice were more susceptible to apoptosis than that of control mice. Stimulating T-cell receptors (TCRs) with anti-CD3 antibody caused upregulation of SERPINB10 expression in polarized Th2 cells, but not polarized Th1 cells. Knockdown of SERPINB10 expression resulted in fewer numbers and greater apoptosis of polarized Th2 cells. Conclusion Our results suggest that SERPINB10 may contribute to allergic inflammation and the Th2 response of asthma by inhibiting the apoptosis of Th2 cells.

scholarly journals Notch ligand Delta-like 4 regulates disease pathogenesis during respiratory viral infections by modulating Th2 cytokines

2007 ◽  
Vol 204 (12) ◽  
pp. 2925-2934 ◽  
Author(s):  
Matthew A. Schaller ◽  
Rupak Neupane ◽  
Brian D. Rudd ◽  
Steven L. Kunkel ◽  
Lara E. Kallal ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Cd4 T Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Interleukin 12 ◽  
Disease Pathogenesis ◽  
Notch Ligand ◽  
Isolated Lung ◽  
Syncytial Virus

Recent data have indicated that an important instructive class of signals regulating the immune response is Notch ligand–mediated activation. Using quantitative polymerase chain reaction, we observed that only Delta-like 4 (dll4) was up-regulated on bone marrow–derived dendritic cells after respiratory syncytial virus (RSV) infection, and that it was dependent on MyD88-mediated pathways. Using a polyclonal antibody specific for dll4, the development of RSV-induced disease was examined. Animals treated with anti-dll4 had substantially increased airway hyperresponsiveness compared with control antibody-treated animals. When the lymphocytic lung infiltrate was examined, a significant increase in total CD4+ T cells and activated (perforin+) CD8+ T cells was observed. Isolated lung CD4+ T cells demonstrated significant increases in Th2-type cytokines and a decrease in interferon γ, demonstrating an association with increased disease pathogenesis. Parellel in vitro studies examining the integrated role of dll4 with interleukin-12 demonstrated that, together, both of these instructive signals direct the immune response toward a more competent, less pathogenic antiviral response. These data demonstrate that dll4-mediated Notch activation is one regulator of antiviral immunity.

Local and Systemic Induction of an Abundant CD4+CD25+ Regulatory T Cell Population by Non-Hodgkin’s Lymphoma.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 357-357
Author(s):  
S. Mittal ◽  
N.A. Marshall ◽  
L. Duncan ◽  
D.J. Culligan ◽  
R.N. Barker ◽  
...  
Keyword(s):  
T Cells ◽  
Tumor Cells ◽  
Peripheral Blood ◽  
Cd4 T Cells ◽  
Treg Cells ◽  
Cell Contact ◽  
Healthy Controls ◽  
Soluble Factors ◽  
Tissue Samples ◽  
Ann Arbor

Abstract Regulatory T (Treg) cells contribute to immune evasion by malignancies. To investigate their importance in non-Hodgkin’s lymphoma (NHL), we enumerated Treg cells in peripheral blood mononuclear cells (PBMC) and involved tissues from 30 newly diagnosed patients. CD25+FoxP3+CD127lowCD4+ Treg cells were increased markedly in PBMC (median=20.4% CD4 T cells, n=20) versus healthy controls (median=3.2%, n=13; p<0. 001, rank sum test) and correlated with serum lactate dehydrogenase (n=14; Rs=0.79, p <0.0001) and disease stage. The median Treg percentage of CD4 T cells from early stages (Ann Arbor stage I and II, n=4) was 12.2%, whereas it was 25.4% in advanced disease (Ann Arbor stages III, IV or bulky stage II, ≥5cm, n=10; p =0.013). We also enumerated Tr1 cells, both in peripheral blood and involved tissue samples, and again compared with healthy controls but no significant differences were noted. We documented poor proliferation of T cells with mitogen ConA and almost none with recall antigens PPD and DPT in both PBMC and involved tissue samples (n=9). T cell hyporesponsiveness was reversed by depleting CD25+ cells (n=4), or by adding anti-CTLA-4 (n=3), supporting the view that Treg cells explain the systemic immunosuppression seen in NHL. A high proportion of Treg cells was also present in involved tissues (median=38.8% CD4 T cells, n=15) versus reactive nodes (median=11.6%, n=2, p=0.02). Therefore, we tested the hypothesis that a regulatory phenotype is induced from conventional T cells within the tumor microenvironment. When autologous CD25- PBMC fractions were incubated with tumor cells from patients (n=6) in vitro, there was consistent strong induction and then expansion of cells with the CD4+CD25+FoxP3+ phenotype of classic ‘natural’ Treg cells as indicated by CFSE dilution. This induction was dependent on tumor dose and was seen when we depleted lymphoid dendritic cells from the involved tissue cell suspension using anti-CD304, or enriched the tumor cells by positive selection of CD20+ cells. This population was confirmed to be suppressive in function (n=3). We also investigated the mechanisms of this induction. Both cell-cell contact and soluble factors appeared important. In two of four cases, some induction was also noted with transwell experiments or with tumor cell conditioned supernatant, indicating that in these cases soluble factors are also involved apart from direct cell-cell contact mechanism. Reports elsewhere suggest roles for prostaglandin E2, tryptophan catabolism, IL-9 and PD-1 interaction with its ligands in inducing a Treg phenotype. Thus, we used cyclooxygenase inhibitors aspirin and sulindac, the indoleamine 2, 3-dioxygenase (IDO) inhibitor 1-methyl tryptophan (1MT), anti-IL-9 receptor antibody and blocking anti-PDL-1 or anti-PDL-2 antibodies in four samples. None of these reagents inhibited Treg induction apart from one case where both anti-PDL-1 and anti-PDL-2 blocking antibodies inhibited Treg induction. We conclude that NHL cells are powerful inducers of Treg cells, which may represent a new therapeutic target.

Expressions of CD3 and CD4 T Cells in Peripheral Blood of Patients with Immunoglobulin A Nephropathy and Their Clinical Significance

2019 ◽  
Vol 9 (6) ◽  
pp. 865-869
Author(s):  
Xuecheng Zhang ◽  
Ning Su ◽  
Dong Chen
Keyword(s):  
T Cells ◽  
Peripheral Blood ◽  
Cd4 T Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Immunoglobulin A ◽  
Immunoglobulin A Nephropathy ◽  
Abnormal Immune Response ◽  
History Of ◽  
Relationship Of ◽  
The Relationship

Immunoglobulin A nephropathy (IgAN) is a primary glomerulonephritis characterized by abnormal immune response-mediated deposition of polymeric IgA (pIgA) in mesangium. As a type of important immune cells, the relationship of CD3 or CD4 with the pathogenesis of IgAN remains poorly understood. In this study, 38 patients with IgAN, 7 patients with idiopathic membranous nephropathy (MN) and 46 healthy adults without history of kidney disease were enrolled. Peripheral blood was collected for further evaluation of the expressions of CD3 and CD4 and IgA by flow cytometry, quantitative polymerase chain reaction (qPCR) and Western blot. Meanwhile, the expression of IgA was detected by ELISA. The result showed that expression of CD3 T cells was down-regulated in patients with IgAN, while amounts of CD4 T cells and IgA level were significantly increased compared to normal control (P < 0.05). However, no signficant changes in CD3, CD4 T cells were found in patients with MN. Our study demonstrates that CD3 and CD4 T cells as well as IgA are involved in the pathogenesis of IgAN and these targets might be beneficial for the treatment of IgAN.

Hypomethylation and Overexpression of CD70 in CD4+ T Cells of the Patients with Immuno-Related Pancytopenia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5157-5157
Author(s):  
Zonghong Shao ◽  
Yue Ren ◽  
Rong Fu
Keyword(s):  
T Cells ◽  
Cd4 T Cells ◽  
Methylation Level ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Quantitative Polymerase Chain Reaction ◽  
Gradient Centrifugation ◽  
Mrna Level ◽  
Venous Blood ◽  
Normal Controls

Abstract Objective To study the expression of CD 70 and the methylation level of CD70 promoter in immuno-related pancytopenia (IRP) patients, and explore the role of CD70 in the pathogenesis of IRP. Methods Thirty-five IRP patients and fifteen healthy donors were enrolled in this study. Peripheral blood mononuclear cells were isolated from venous blood by density gradient centrifugation and CD4+ T cells were isolated by immunomagnetic beads.The mRNA level and the percentage of CD70 of CD4+ T cells were measured by real-time quantitative polymerase chain reaction (RT-PCR) and flow cytometry respectively. Methylation-Specific PCR (MSP) was performed to determine the methylation level of CD70 promoter. Results The percentage of CD70 expression on CD4+ T cells of untreated IRP patients[(7.46±1.51)%] was significantly higher than that of recovered IRP patients [(5.95±1.34) %] and normal controls [(1.83±0.6)%] and the result of recovered IRP patients was significantly higher than that of normal controls (P<0.05).The relative expressions of CD70 mRNA in CD4+ T cells were [2.314(0.200~6.084)]A[1.021 (0.135~3.434)]A [0.353 (0.008~2.258)] in three groups respectively. The differences between untreated IRP patients,recovered IRP patients and normal controls were significant (P<0.05). The CD70 promoter methylation level in CD4+ T cells of all IRP patients was significantly lower than that of normal controls (p<0.05). The expression of CD70 positively correlated to the ratio of CD5+ B cells (r=0.533, p<0.001). Conclusions The overexpression of CD70 may lead to immunologic disarrangement and patients and it may play important role in the pathogenesis of IRP. Disclosures No relevant conflicts of interest to declare.

scholarly journals SERPINB10 Contributes to Asthma by Inhibiting the Apoptosis of Allergenic Th2 Cells

2020 ◽  
Author(s):  
Yuqing Mo ◽  
Ling Ye ◽  
Hui Cai ◽  
Guiping Zhu ◽  
Jian Wang ◽  
...  
Keyword(s):  
T Cells ◽  
Allergic Asthma ◽  
Cd4 T Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Allergic Inflammation ◽  
Specific Ige ◽  
Th2 Cells ◽  
Th1 Cells ◽  
Th2 Response ◽  
Th1 And Th2

Abstract Background: Serine peptidase inhibitor, clade B, member 10 (SERPINB10) contributes to allergic inflammation in asthma. However, its role in the T-helper type 1 (Th1) /Th2 response of allergic asthma is not known. The goal of this study was to unveil the function of SERPINB10 in the Th2 response of allergic asthma and the mechanism by which SERPINB10 affects the viability of Th2 cells.Methods: Th2 cytokines and serum levels of house dust mite (HDM)-specific IgE in bronchoalveolar lavage fluid were examined by ELISA in an HDM-induced asthma model. The number and apoptosis of Th1 and Th2 cells in mouse lungs were measured by flow cytometry. Naïve CD4 T cells from patients with asthma were cultured under appropriate polarizing conditions to generate Th1 and Th2 cells. SERPINB10 expression in polarized Th1 and Th2 cells was quantified by real-time reverse transcription-quantitative polymerase chain reaction. SERPINB10 expression was knocked down in CD4 T cells with lentivirus. Results: Knockdown of SERPINB10 expression significantly diminished HDM-induced Th2 cytokine secretion and level of HDM-specific IgE. After HDM exposure, SERPINB10-knockdown mice had diminished numbers of Th2 cells, but similar numbers of Th1 cells, compared with those in negative-control mice. Th2 cells of SERPINB10-knockdown mice were more susceptible to apoptosis than that of control mice. Stimulating T-cell receptors (TCRs) with anti-CD3 antibody caused upregulation of SERPINB10 expression in polarized Th2 cells, but not polarized Th1 cells. Knockdown of SERPINB10 expression resulted in fewer numbers and greater apoptosis of polarized Th2 cells.Conclusion: Our results suggest that SERPINB10 may contributes to allergic inflammation and the Th2 response of asthma by inhibiting the apoptosis of Th2 cells.

scholarly journals The effects of acute oxygen changes on heart rate in the freshwater crab Poppiana dentata (Randall, 1840)

2020 ◽  
Vol 48 (3) ◽  
pp. 480-487
Author(s):  
Delezia Shivani Singh ◽  
Mary Alkins-Koo ◽  
Luke Victor Rostant ◽  
Azad Mohammed
Keyword(s):  
Heart Rate ◽  
Invasive Technique ◽  
Cardiac Physiology ◽  
Low Oxygen ◽  
Heart Rates ◽  
Oxygen Levels ◽  
Non Invasive ◽  
Hypoxic Conditions ◽  
Cardiac Responses ◽  
Insight Into

Heart rate is a key physiological feature that can be used to assess the response of organisms to changing environmental conditions in aquatic habitats, such as acute fluctuations in oxygen levels and hypoxic conditions. This experiment, therefore, investigated cardiac responses in a freshwater brachyuran species, Poppiana dentata, exposed to low oxygen levels. Heart rate was derived from beats per minute (bpm) signals (n = 576) using an infrared, non-invasive technique over a 96 h period, under different dissolved oxygen (DO) conditions. These involved three regimes: normoxic (6.8 ± 0.1 mg L-1), decreasing DO to hypoxic levels (6.2 to 1.7 mg L-1), and recovery with normoxic levels (6.3 ± 0.1 mg L-1). Changes in heart rates among the three regimes were significant (P < 0.05); reflecting the shift in heart rate during different conditions of oxygen availability, normoxic (59 to 61 bpm), declining DO (54 to 62 bpm) and recovery DO (53 to 64 bpm). Additionally, the normal rhythmicity of heart rates under the normoxic condition was not maintained throughout most of the declining DO and recovery periods. P. dentata has demonstrated cardiac compensations in heart rate during low oxygen levels, providing insight into the species cardiac physiology.

scholarly journals AB0075 SYNOVIAL TISSUE MACROPHAGES ARE DOMINANTLY ALTERNATIVELY ACTIVATED IN PATIENTS WITH MATURE OSTEOARTHRITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1337.2-1338
Author(s):  
G. Laskarin ◽  
T. Kehler ◽  
D. Legović ◽  
V. Šantić ◽  
B. Ćurko-Cofek ◽  
...  
Keyword(s):  
T Cells ◽  
Knee Osteoarthritis ◽  
Synovial Tissue ◽  
Tnf Alpha ◽  
Lymphocyte Infiltration ◽  
Activated Macrophages ◽  
Tissue Samples ◽  
Arginase 1 ◽  
Immunofluorescence Labeling ◽  
Alternatively Activated

Background:Macrophages are abundant inflammatory cell type in the synovial membrane of knee osteoarthritis (OA) (1). Their quantity is associated with radiographic severity of knee OA and joint symptoms (2), while their functions are set in response to micro-environmental signals (3). Classically activated macrophages M1 support T helper 1 (Th1) driven pro-inflammatory reactions, while alternatively activated macrophages M2 strengthen Th2 inflammatory processes (3).Objectives:To investigate activation status of synovial tissue macrophages in patients with mature OA in terms of M1 / M2 polarization.Methods:Synovial tissue samples (6) with abundant lymphocyte infiltration were obtained during aloarthroplasty. Double immunofluorescence labeling was performed on paraffin-embedded synovial tissue sections using primary rabbit anti-macrophage CD68 mAb in combination with mouse anti-human antibodies directed toward CD3, arginase-1, TNF-alpha and IL-15. CD206 and CD163 were single labelled.Results:CD68+ macrophages mostly co-expressed arginase-1 (4/6 samples), indicating their M2 orientation. Macrophages were placed in lining synovial tissue and nearby tissue-resident CD3+ cells. M2 markers CD206 and CD163 were found in the area of macrophage interaction with T cells. CD68+ cells co-expressing TNF-alpha or IL-15 M1 markers were in minority in these synovial tissues. Lymphocyte infiltration was less abundant in remaining (2/6) synovial tissue samples.Conclusion:Mature synovial tissue macrophages, equipped dominantly with arginase-1 are M2 oriented and might support Th2 immune response in surrounding T cells.References:[1]Grieshaber-Bouyer R, Kämmerer T, Rosshirt N, Nees TA, Koniezke P, Tripel E, Schiltenwolf M, Kirsch J, Hagmann S, Moradi B. Divergent Mononuclear Cell Participation and Cytokine Release Profiles Define Hip and Knee Osteoarthritis. J Clin Med. 2019;8(10):piiE1631.[2]Haraden CA, Huebner JL, Hsueh MF, Li YJ, Kraus VB. Synovial fluid biomarkers associated with osteoarthritis severity reflect macrophage and neutrophil related inflammation. Arthritis Res Ther. 2019;21(1):146.[3]Barros MH, Hauck F, Dreyer JH, Kempkes B, Niedobitek G. Macrophage polarisation: an immunohistochemical approach for identifying M1 and M2 macrophages. PLoS One. 2013;8(11):e80908.Acknowledgments:University of Rijeka supported the research by the grants No. Uni-ri-biomed-18-110 and No. Uni-ri-biomed-18-160.Disclosure of Interests:None declared

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