Figure 6. Eomes is critical for Vγ4 γδ T cell IFN-γ production.

(A) CD44^high Vγ4 and Vγ1 γδ T cells were sorted from B6 mice for real-time PCR analysis. T-bet and Eomes transcripts were normalized against HPRT. One example of 3 repeated experiments is shown. (B). Vγ1 or Vγ4 cells were expanded as described above and the transcription of T-bet and Eomes was determined by real-time PCR. (C). Naïve (CD44^low) Vγ4 and Vγ1 γδ T cells were sorted from wild-type mice and activated with anti-CD3 and anti-CD28 in the presence of IL-12 and anti-IL-4. Cells were then infected with either control, DN T-bet or DN Eomes retrovirus-GFP as described in Materials and Methods. After 48hrs, cells were cultured in the presence of IL-2 for additional 3 days, restimulated with anti-CD3 and anti-CD28 for 6h for intracellular cytokine staining. One representative experiment for the percentage of IFN-γ⁺ reduction ((% of IFN-γ⁺ cells from control retrovirus infected - % of IFN-γ⁺ cells from DN retrovirus infected)/ % of IFN-γ⁺ cells from control retrovirus infected) is shown.