Figure 4. GVHD increases apoptosis 7 days after adoptive transfer of TCRTg CD8⁺, but not CD4⁺, T cells.

Mice were transplanted per Figure 1A and given either an infusion of Marilyn or Matahari splenocytes on day +28 with or without a male B6 DC vaccine. Apoptosis of nonproliferated, CFSE⁺, TCRTg T cells was enumerated by flow cytometric analysis of Annexin V at 3, 5 and 7 days (days +31, 33 and 35 post-BMT) after transfer of CD4⁺ Marilyn T cells in (A) unvaccinated and (B) male DC vaccinated recipient lymph nodes, and after transfer of CD8⁺ Matahari T cells in (C) unvaccinated and (D) male DC vaccinated recipient lymph nodes, 3-7 mice/group, pooled from three independent experiments. (E) Matahari T cells were cultured with male or female DCs from either allogeneic (C3H.SW) or syngeneic (B6 CD45.1⁺) donors at a 1:1 ratio for 72 hours. Cells were then counted and analyzed for expression of Annexin V on CD45.2⁺CD8⁺Vβ 8.3⁺7AAD^- cells. Triplicate co-culture wells were set up for each condition. * p<0.05.