Figure 1.

CD27 expression defines distinct subsets of peripheral γδ T cells. (a) Flow cytometry of cells obtained from various tissues (above plots) of adult wild- type C57BL/6 mice, stained for CD3ε, CD27 and TCRγδ and gated on CD3ε⁺TCRγδ⁺ cells. a+i(LN), axillary and inguinal lymph nodes; mLN, mesenteric lymph nodes. Data are representative of three to eight independent experiments with three to five mice each. (b) Flow cytometry of splenocytes stained for CD3ε, CD27 and TCRγδ, plus CD62L or CD44 and gated on CD3ε⁺TCRγδ⁺CD27⁺ cells (γδ²⁷⁺) and CD3ε⁺TCRγδ⁺CD27⁻cells (γδ²⁷⁻). Data are representative of three independent experiments with three to five mice each. (c) Flow cytometry of γδ²⁷⁺ or γδ²⁷⁻ T cells sorted from pooled spleens and lymph nodes and cultured separately for 3 d with antigen-presenting cells and anti-CD3ε (1 mg/ml; left) or injected separately intravenously into RAG-2- deficient mice and analyzed after 2 weeks (right), then recovered, stained for CD3ε, CD27 and TCRδ and gated on CD3ε⁺TCRδ⁺ cells. Data are representative of three independent experiments. (d–f) Flow cytometry analyzing the proliferation of γδ²⁷⁺ or γδ²⁷⁻ T cells sorted from pooled spleens and lymph nodes, labeled with CFSE and cultured separately with antigen- presenting cells in presence of various doses of anti-CD3e (horizontal axis; d) or injected intravenously into RAG-2-deficient mice separately (e) or together at ratio of 1:1 (f), and assessed on the basis of CFSE-dilution kinetics at day 3. Data are representative of four independent experiments.