scholarly journals Ex Vivo Characterization of Early Secretory Antigenic Target 6-Specific T Cells at Sites of Active Disease in Pleural Tuberculosis

2005 ◽  
Vol 40 (1) ◽  
pp. 184-187 ◽  
Author(s):  
K. A. Wilkinson ◽  
R. J. Wilkinson ◽  
A. Pathan ◽  
K. Ewer ◽  
M. Prakash ◽  
...  
Keyword(s):  
T Cells ◽  
Ex Vivo ◽  
Pleural Tuberculosis ◽  
Early Secretory Antigenic Target ◽  
Active Disease

scholarly journals Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood

2001 ◽  
Vol 193 (11) ◽  
pp. 1303-1310 ◽  
Author(s):  
Detlef Dieckmann ◽  
Heidi Plottner ◽  
Susanne Berchtold ◽  
Thomas Berger ◽  
Gerold Schuler
Keyword(s):  
T Cells ◽  
Cytotoxic T Lymphocyte ◽  
Ex Vivo ◽  
Cell Receptor ◽  
Regulatory Cells ◽  
Suppressor T Cells ◽  
Isolation And Characterization ◽  
Regulatory Properties

It has been known for years that rodents harbor a unique population of CD4+CD25+ “professional” regulatory/suppressor T cells that is crucial for the prevention of spontaneous autoimmune diseases. Here we demonstrate that CD4+CD25+CD45RO+ T cells (mean 6% of CD4+ T cells) are present in the blood of adult healthy volunteers. In contrast to previous reports, these CD4+CD25+ T cells do not constitute conventional memory cells but rather regulatory cells exhibiting properties identical to their rodent counterparts. Cytotoxic T lymphocyte–associated antigen (CTLA)-4 (CD152), for example, which is essential for the in vivo suppressive activity of CD4+CD25+ T cells, was constitutively expressed, and remained strongly upregulated after stimulation. The cells were nonproliferative to stimulation via their T cell receptor for antigen, but the anergic state was partially reversed by interleukin (IL)-2 and IL-15. Upon stimulation with allogeneic (but not syngeneic) mature dendritic cells or platebound anti-CD3 plus anti-CD28 the CD4+CD25+ T cells released IL-10, and in coculture experiments suppressed the activation and proliferation of CD4+ and CD8+ T cells. Suppression proved IL-10 independent, yet contact dependent as in the mouse. The identification of regulatory CD4+CD25+ T cells has important implications for the study of tolerance in man, notably in the context of autoimmunity, transplantation, and cancer.

scholarly journals Ex vivo purification and characterization of human invariant natural killer T cells

2011 ◽  
Vol 373 (1-2) ◽  
pp. 1-7 ◽  
Author(s):  
Ashish Arunkumar Sharma ◽  
Leon Chew ◽  
Mihoko Ladd ◽  
Roger Jen ◽  
Pascal M. Lavoie
Keyword(s):  
T Cells ◽  
Natural Killer ◽  
Ex Vivo ◽  
Natural Killer T Cells ◽  
Purification And Characterization ◽  
Killer T Cells ◽  
Invariant Natural Killer ◽  
Natural Killer T

scholarly journals Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

2020 ◽  
Author(s):  
Christina Gerstner ◽  
Sara Turcinov ◽  
Aase H Hensvold ◽  
Karine Chemin ◽  
Hannes Uchtenhagen ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
At Risk ◽  
T Cells ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Class Ii ◽  
Hla Class Ii ◽  
Phenotypic Characterization ◽  
Early Ra

Abstract Background: HLA class II tetramers can be used for ex vivo enumeration and phenotypic characterization of antigen-specific CD4+ T cells. They are increasingly applied in settings like allergy, vaccination and autoimmune diseases. Rheumatoid arthritis (RA) is a chronic autoimmune disorder for which many autoantigens have been described. Results: Using multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificities in RA patient samples. We focused on previously described citrullinated HLA-DRB1*04:01-restricted T cell epitopes from α-enolase, fibrinogen-b, vimentin as well as cartilage intermediate layer protein (CILP).First, we examined inter-assay variability and the sensitivity of the assay in peripheral blood from healthy donors (n=7). Next, we confirmed the robustness and sensitivity in a cohort of RA patients with repeat blood draws (n=14). We then applied our method in two different settings. We assessed lymphoid tissue from seropositive arthralgia (n=5) and early RA patients (n=5) and could demonstrate autoreactive T cells in individuals at risk of developing RA. Lastly, we studied peripheral blood from early RA patients (n=10) and found that the group of patients achieving minimum disease activity (DAS28 <2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific T cells. Conclusions: Our study demonstrates the development of a sensitive tetramer panel allowing simultaneous characterization of antigen-specific T cells in ex vivo patient samples including RA ‘at risk’ subjects. This multi-tetramer approach can be useful for longitudinal immune-monitoring in any disease with known HLA-restriction element and several candidate antigens.

scholarly journals Integrative molecular profiling of autoreactive CD4 T cells in autoimmune hepatitis

2020 ◽  
Author(s):  
Amédée Renand ◽  
Iñaki Cervera-Marzal ◽  
Laurine Gil ◽  
Chuang Dong ◽  
Erwan Kervagoret ◽  
...  
Keyword(s):  
T Cells ◽  
Disease Activity ◽  
Autoimmune Hepatitis ◽  
Peripheral Blood ◽  
Cd4 T Cells ◽  
Ex Vivo ◽  
B Cell Differentiation ◽  
Tcr Repertoire ◽  
First Time

AbstractBackground & AimsIn most autoimmune disorders, crosstalk of B cells and CD4 T cells results in the accumulation of autoantibodies. In autoimmune hepatitis (AIH), the presence of anti-Soluble Liver Antigen (SLA or SepSecs) autoantibodies is associated with significantly reduced overall survival, but the associated autoreactive CD4 T cells have not been characterized yet. Here we isolated and deeply characterized SLA-specific CD4 T cells in AIH patients.MethodsWe used brief ex vivo restimulation with overlapping SLA-derived peptides to isolate and phenotype circulating SLA-specific CD4 T cells, and integrative single-cell RNA-seq (scRNA-seq) to characterize their transcriptome and TCR repertoire in n=5 AIH patients. SLA-specific CD4 T cells were tracked in peripheral blood through TCR sequencing, to identify their phenotypic niche. We further characterized disease-associated peripheral blood T cells by high content flow cytometry in an additional cohort of n=46 AIH patients and n=18 non-alcoholic steatohepatitis (NASH) controls.ResultsAutoreactive SLA-specific CD4 T cells were only detected in patients with anti-SLA autoantibodies and had a memory PD-1+CXCR5−CCR6−CD27+ phenotype. ScRNA-seq revealed their pro-inflammatory/B-Helper profile (IL21, IFNG, TIGIT, CTLA4, NR3C1, CD109, KLRB1 and CLEC2D). Autoreactive TCR clonotypes were restricted to the memory PD-1+CXCR5− CD4 T cells. This subset was significantly increased in the blood of AIH patients and supported B cell differentiation through IL-21. Finally, we identified a specific phenotype (PD-1+CD38+CD27+CD127−CXCR5−) of CD4 T cells linked to disease activity and IgG response during AIH.ConclusionsThis work provides for the first time a deep characterization of rare circulating autoreactive CD4 T cells and the identification of their peripheral reservoir in AIH. We also propose a generic phenotype of pathogenic CD4 T cells related to AIH disease activity.

Ex vivo characterization of γδ T-cell repertoire in patients after adoptive transfer of Vγ9Vδ2 T cells expressing the interleukin-2 receptor β-chain and the common γ-chain

Cytotherapy ◽  
2013 ◽  
Vol 15 (4) ◽  
pp. 481-491 ◽  
Author(s):  
Takamichi Izumi ◽  
Makoto Kondo ◽  
Takuya Takahashi ◽  
Nao Fujieda ◽  
Atsushi Kondo ◽  
...  
Keyword(s):  
T Cells ◽  
Ex Vivo ◽  
Interleukin 2 ◽  
T Cell Repertoire ◽  
Γδ T Cell ◽  
Cell Repertoire ◽  
Vγ9vδ2 T Cells ◽  
The Common ◽  
Β Chain

scholarly journals Ex-vivo characterization of regulatory T cells in pulmonary tuberculosis patients, latently infected persons, and healthy endemic controls

Tuberculosis ◽  
2016 ◽  
Vol 100 ◽  
pp. 61-68 ◽  
Author(s):  
Martha Zewdie ◽  
Rawleigh Howe ◽  
Søren T. Hoff ◽  
T. Mark Doherty ◽  
Nahom Getachew ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Pulmonary Tuberculosis ◽  
Ex Vivo ◽  
Tuberculosis Patients ◽  
Latently Infected

scholarly journals Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

2020 ◽  
Author(s):  
Christina Gerstner ◽  
Sara Turcinov ◽  
Aase H Hensvold ◽  
Karine Chemin ◽  
Hannes Uchtenhagen ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
At Risk ◽  
T Cells ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Class Ii ◽  
Hla Class Ii ◽  
Phenotypic Characterization ◽  
Early Ra

Abstract Background: HLA class II tetramers can be used for ex vivo enumeration and phenotypic characterization of antigen-specific CD4+ T cells. They are increasingly applied in settings like allergy, vaccination and autoimmune diseases. Rheumatoid arthritis (RA) is a chronic autoimmune disorder for which many autoantigens have been described.Results: Using multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificities in RA patient samples. We focused on previously described citrullinated HLA-DRB1*04:01-restricted T cell epitopes from α-enolase, fibrinogen-b, vimentin as well as cartilage intermediate layer protein (CILP). First, we examined inter-assay variability and the sensitivity of the assay in peripheral blood from healthy donors (n=7). Next, we confirmed the robustness and sensitivity in a cohort of RA patients with repeat blood draws (n=14). We then applied our method in two different settings. We assessed lymphoid tissue from seropositive arthralgia (n=5) and early RA patients (n=5) and could demonstrate autoreactive T cells in individuals at risk of developing RA. Lastly, we studied peripheral blood from early RA patients (n=10) and found that the group of patients achieving minimum disease activity (DAS28 <2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific T cells. Conclusions: Our study demonstrates the development of a sensitive tetramer panel allowing simultaneous characterization of antigen-specific T cells in ex vivo patient samples including RA ‘at risk’ subjects. This multi-tetramer approach can be useful for longitudinal immune-monitoring in any disease with known HLA-restriction element and several candidate antigens.

scholarly journals Establishment and Characterization of a Cell Based Artificial Antigen-Presenting Cell for Expansion and Activation of CD8+ T Cells Ex Vivo

2008 ◽  
Vol 5 (1) ◽  
pp. 47-53 ◽  
Author(s):  
Weijuan Gong ◽  
Mingchun Ji ◽  
Zhengfeng Cao ◽  
Liheng Wang ◽  
Yayun Qian ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Ex Vivo ◽  
Antigen Presenting Cell ◽  
Artificial Antigen ◽  
A Cell ◽  
Antigen Presenting
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