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Updated: Aug 11, 2026
Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry
Published on: January 7, 2019
[Isolation, identification and functional characterization of human CD4+ CD25+ Treg cells from human peripheral
Jiang-hua Yang1, Yong-xiang Zhang, Fang Wang
1Department of Infectious Diseases, the First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, China. yjhpath@163.com
Aim:
To isolate CD4(+) CD25(+) Treg cells from human peripheral blood, and study their functional characteristics.
Methods:
The expressions of Foxp3 in human CD4(+) CD25(+) Treg cells were test by RT-PCR. The regulatory properties of CD4(+) CD25(+) Treg cells were assessed by co-culturing with CD4(+) CD25(-) T cells and CD8(+) T cells, or adding exogenous IL-2. The detection of intracellular cytokine production of IL-4, IL-10 and IFN-gamma was done by flow cytometry.
Results:
CD4(+) CD25(+) Treg cells highly expressed Foxp3 and mainly synthesized IL-10 that suppressed the proliferation of CD4(+) CD25(-) T and CD8(+) T cells. Its suppressive function was reversed by high concentration of IL-2 and (or) IL-4.
Conclusion:
CD4(+) CD25(+) Treg cells exhibited a subpopulation of immunoregulatory T cells with suppressive function, which could be reversed by high concentration of IL-2.
