Figure 1.

Superior HIV replication in Th1Th17 vs. Th1 cells is regulated at entry and post-entry level. Matched Th1 and Th1Th17 cells were sorted by flow cytometry from four different HIV-uninfected subjects and stimulated via CD3/CD28 for 3 days (A). Cells were exposed to replication competent NL4.3BAL-GFP (B) or single round HIV-VSVG-GFP pseudotyped strains (50 ng HIV-p24/10⁶ cells) (C-D) for 3 h at 37°C. Unbound virus was removed by extensive washing, and cells were cultured for 3 additional days in the presence of IL-2 (5 ng/ml). Integrated HIV-DNA and GFP expression levels were quantified by nested real-time PCR (B-C) and flow cytometry (D), respectively, at day 3 post-infection. Shown in B-C is relative HIV-DNA integration in Th1 vs. Th1Th17 cells (normalized to the maximal value considered to be 100% in Th1Th17 cells); values above graphs are integrated HIV-DNA copies per 10⁶ cells in Th1Th17 cells (mean±SD of triplicate wells). Paired t-Test values are indicated on the graphs.