Figure 40.

Phenotyping of Treg cells from murine spleen and lymph nodes. (A) Gating strategy to identify Treg cells in the spleen. From all events, lymphocytes can be distinguished by their FSC/SSC properties (gate G0). Based on G0, doublets are excluded twice (gates G1 and G2) followed by exclusion of dead or autofluorescent cells (gate G3). From G3, CD4⁺CD3ε⁺ T cells (gate G4) are gated, from which Foxp3⁺ Treg cells (gate G6) and Foxp3⁻ Tconv cells (gate G5) can be further identified. From G6, Helios⁺ tTreg (gate G7) and Helios⁻ pTreg cells (gate G8) are gated. Finally, a staining for CD62L and CD44 on Treg cells (gate G6, blue) and Tconv cells (gate G5, orange) are shown together, with CD62L⁻CD44⁺ effector/memory cells being gated (gate G9). (B and C) Gating strategy to identify Treg cells in skin-draining lymph nodes (B) and mesenteric lymph nodes (C). Gates as described in panel A. Numbers indicate frequencies of cells within respective gates. Figures are based on spleen and lymph node isolations from wild-type mice.