HIV p24Gag-specific T cell cytokine responses in spleens of uninfected or Mtb-infected mice vaccinated with MultiHIV DNA/protein.

Splenocytes from Mtb-infected or uninfected mice, which were immunized and boosted with MultiHIV DNA/protein, were isolated 4 wk post-vaccination, restimulated ex vivo with p24Gag antigen, and stained for CD4 and CD8 markers and for intracellular IFN-γ, TNF, and IL-2 (details in Materials and Methods). Numbers of p24Gag-specific CD4+ (A) and CD8+ (B) T cells producing one of three cytokines were determined by flow cytometry. Mean number of single cytokine-producing T cells ± SEM are shown (6–8 animals/group from one individual experiment; **: P<0.01). Flow cytometry plots depict representative intracellular cytokine staining of CD4+ (C) and CD8+ (D) T cells. The cytokine profile of p24Gag-specific T cells was determined in two separate experiments.