Fig. 2. Deficient activation of CD4⁺ T cells from BM chimeric mice reconstituted with PKCθ knockout BM cells expressing PKCθ^HR2A.

(A–D) Flow cytometry analysis of the surface abundance of CD25 and CD69 (A and B) and the production of IL-2 (C and D) by transduced (GFP⁺) or non-transduced (GFP⁻) naïve CD4⁺ T cells from BM chimeric mice reconstituted with PKCθ knockout BM cells expressing empty vector (vector), PKCθ^WT, or PKCθ^HR2A. Cells were left unstimulated or restimulated with CD3 and CD28 mAbs. Numbers above bracketed lines (A) and in the gated boxes (C) indicate % positive cells from a single representative mouse in each group. MFI values of CD25 and CD69 (A) or IL-2 (C) are shown, and averaged data pooled from six mice/group are shown in (B) and (D). *P < 0.05, **P < 0.01, ***P < 0.001. Results are representative of 4 independent experiments.

(E and F) Flow cytometry analysis of the proliferation (E) and the amounts of IFNγ (Th1) or IL-4 (Th2) cytokines (F) in sorted (GFP⁺) CD4⁺ cells as in (A to D). Numbers in (E) and (F) indicate the % of proliferating cells or the frequency of IFNγ- or IL-4-containing cells among gated CD4⁺ cells, respectively. Results are representative of 6 mice/group and 3 independent experiments.