Figure 3. Vaccine-induced proliferation is impaired for TCRTg CD4⁺ and CD8⁺ T cells during GVHD.

Mice were transplanted per Figure 1A and received CFSE-labeled CD4⁺ Marilyn or CD8⁺ Matahari splenocytes with or without a male B6 DC vaccine. Proliferation was measured by flow cytometry at 3, 5 and 7 days (days +31, 33 and 35 post-BMT) after transfer. (A) Results from a representative recipient of CD4⁺ Marilyn T cells after allogeneic and syngeneic BMT analyzed at each timepoint are depicted, with unvaccinated recipient (top), and male DC vaccinated recipient (bottom). (B) Pooled data from three independent experiments depicted for CD4⁺ Marilyn T cells in unvaccinated, and (C) male DC vaccinated recipients, as well as from three independent experiments for (D) CD8⁺ Matahari T cells in unvaccinated, and (E) male DC vaccinated recipients, 3-9 mice/group. CD8⁺ Matahari T cells from syngeneic and allogeneic recipients were analyzed 3 days following vaccination with male DC vaccine for intracellular IFNγ (F) or CD107a (G) (n=3 mice/group). (H) Representative FACS plots of PD-1 and TIM3 on CD4⁺ Marilyn T cells from syngeneic and allogeneic recipients 3 days after male DC vaccination (total of n=3 mice per group analyzed). * p<0.05, ** p<0.01, *** p<0.001.