Figure 8.

Bcl11b expression is critical for the magnitude and functionality of primary effector CD8 T cells after respiratory infection with VacV. WT (Bcl11b^flox/flox/dLck-iCre⁻; n = 8) or Bcl11b-conditional knockout (Bcl11b^flox/flox/dLck-iCre⁺; n = 8) mice were infected i.n. with VacV-WR (1.5 × 10⁴ PFU/mouse). Eight days postinfection, splenocytes and lung cells were harvested and stained for CD8, CD44, Ki67, and B8R_20–27/k^b tetramer. (A,B) Left, representative plots of CD8/CD44 (Top Panels) and CD8⁺CD44^hi B8R_20–27/k^b-tetramer staining (Bottom Panels), gating on live cells, are shown. Percentages of activated (CD44^hi) and B8R_20–27/k^b tetramer + CD8 T cells within each gate are indicated. Right, percentages and total numbers of CD8⁺CD44^hi and B8R_20–27/k^b tetramer + cells per spleen (A) and lung (B). (C) Representative plots for intracellular IFN-γ staining in gated CD8+CD44+ T cells. Settings were based on controls, after gating on naïve CD44^lo cells in the same host. Percentages that stained positive for each marker are indicated. (D) Left, representative plots showing the percentage of CD8 T cell proliferation by Ki67 staining among CD44^hi cells in VacV-infected mice. Right, percentages and total numbers of CD8⁺CD44^hiKi67⁺ cells per spleen and lung. Settings were based on controls, after gating on naïve CD44^lo cells in the same host. Percentages that stained positive for each marker are indicated. The results shown are representative of two separate experiments each with four mice per group. Asterisks indicate statistical significance. The p-values are <0.05 by two-tailed Student t-test (WT vs. Cko).