A rapid diagnostic test for human regulatory T-cell function to enable regulatory T-cell therapy.
Regulatory T cells (CD4(+)CD25(hi)CD127(lo)FOXP3(+) T cells [Tregs]) are a population of lymphocytes involved in the maintenance of self-tolerance. Abnormalities in function or number of Tregs are a feature of autoimmune diseases in humans. The ability to expand functional Tregs ex vivo makes them ideal candidates for autologous cell therapy to treat human autoimmune diseases and to induce tolerance to transplants. Current tests of Treg function typically take up to 120 hours, a kinetic disadvantage as clinical trials of Tregs will be critically dependent on the availability of rapid diagnostic tests before infusion into humans. Here we evaluate a 7-hour flow cytometric assay for assessing Treg function, using suppression of the activation markers CD69 and CD154 on responder T cells (CD4(+)CD25(-) [Tresp]), compared with traditional assays involving inhibition of CFSE dilution and cytokine production. In both freshly isolated and ex vivo expanded Tregs, we describe excellent correlation with gold standard suppressor cell assays. We propose that the kinetic advantage of the new assay may place it as the preferred rapid diagnostic test for the evaluation of Treg function in forthcoming clinical trials of cell therapy, enabling the translation of the large body of preclinical data into potentially useful treatments for human diseases.
Medical Research Council Centre for Transplantation, King's College London, United Kingdom.
James B
JB
Canavan
Behdad
B
Afzali
Cristiano
C
Scottà
Henrieta
H
Fazekasova
Francis C
FC
Edozie
Thomas T
TT
Macdonald
Maria P
MP
Hernandez-Fuentes
Giovanna
G
Lombardi
Graham M
GM
Lord
D015703:Antigens, CD / Q000276:immunology* / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology* / Q000378:metabolism; D001327:Autoimmune Diseases / Q000276:immunology / Q000628:therapy; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D023201:CD40 Ligand / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D005452:Fluoresceins; D006801:Humans; D007108:Immune Tolerance / Q000276:immunology; D007159:Immunologic Tests / Q000379:methods*; D037181:Lectins, C-Type / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results; D013388:Succinimides; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D013997:Time Factors; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D023201:CD40 Ligand; D016207:Cytokines; D005452:Fluoresceins; D037181:Lectins, C-Type; D013388:Succinimides; D000099192:CD69 Antigens; C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester