Figure 6.
Differential HIV-specific CD4+ T cell and CD8+ T cell cytokine response. (A) Stimulation of patient PBMCs cells isolated directly ex vivo with overlapping Gag peptides. The percent of IFN-γ expressing HIV-specific CD4+ T cells that co-expressed IL-2 is shown for VC1 and CP2. A minimum of 250,000 total events were analyzed by FACS. (B) Representative FACS plots showing the stimulation of PBMCs isolated from VC1 and CP1. The plots are gated on all CD8+ T cells, and the percentage of IFN-γ positive cells that co-expressed TNF-α or perforin are indicated in the top right quadrant. Quantification of the percentage of IFN-γ expressing CD8+ T cells that co-expressed perforin, TNF-α, or both perforin and TNF-α after stimulation with patient-specific Gag (C) or Nef (D) peptides are shown for both VC1 and CP1. A minimum of 250,000 total events were analyzed by FACS. Comparative CD8+ T cell inhibition of viral replication. (E) CD4+ T cells from VC1 and CP1 were infected with GFP expressing recombinant NL4-3 virus. CD4+ T cells were cultured alone or with autologous CD8+ T cells at a 1:1 ratio. Percent infection was measure five days after infection by FACS analysis. (F) CD4+ T cells from 4 Healthy Donors (D1–4), an untreated Viremic patient, 6 ES, and VC1 were infected with GFP expressing replication-competent NL4-3 virus. CD4+ T cells were cultured alone or with autologous CD8+ T cells at a 1:1 ratio. Percent inhibition was quantified 5 days after infection.