A flow cytometry based assay for the enumeration of regulatory T cells in whole blood.

摘要: The analysis of regulatory T cells (T-reg(s)) is becoming an increasingly important consideration in the development novel immunotherapeutic strategies. Accurate quantification T-regs during treatment protocols crucial, particularly where therapeutic strategy targeting T-regs. TruCOUNT™ method has utility for enumerating different immune but not been used to detect We have utilized this technology develop assay enumerate human whole blood, based on CD127 expression. mean number CD4+CD25+CD127lo per μl blood was 48±16.9 with a range 18 - 79 (n=22) and average percentage 6.1±1.9% (range 2.2-10.4%). percentages were similar when detected or density-gradient separated PBMC, comparable those distinguished using T-reg marker FoxP3. robust reliable enumeration lower frequency T-regs, CV's intra-assay repeatability inter-assay precision <9% <35%, respectively. detection total CD4+ lymphocytes <2% <18% precision, providing further evidence reproducibility. This advantages over current methods, including small sample volume, ability determine absolute cell counts, no need hematology analyzers. will simplify clinical trial monitoring can be provide crucial data patient numbers before, during, after interventions.