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. Author manuscript; available in PMC: 2020 Jan 3.
Published in final edited form as: J Immunol. 2018 Nov 2;201(11):3269–3281. doi: 10.4049/jimmunol.1800940

FIGURE 1.

FIGURE 1.

Flow cytometry gating strategy for Treg cell analysis. PBMCs were gated first on viability and on CD3 and CD4 positivity. (A–D) (A) CD4+ Treg cells were defined by high expression of CD25, and intracellular FOXP3 and conventional CD4+ T cells (Th CD4) were defined by low expression of CD25 and FOXP3. (B) Treg cells were then analyzed for CD45RA and CD27 expression, and populations were defined by dual or single expression. (C–F) CLA and CCR4 expression was also analyzed among Treg cells. CLA- and CCR4-positive Treg cells were also confirmed to be CD45RA negative with mixed expression of CD27. (G–I) (E) Six groups of CD4+ T cells were defined by CD45RA positivity and various CD25 expression. (H) Each population was analyzed for FOXP3 expression. (I) Distribution of these populations was analyzed in CD45RA−/CD27+ (Central) Treg cells, CD45RA−/CD27− (Effector) Treg cells, CLA+ Treg cells, and CCR4hi Treg cells as defined in (B)–(D).