Figure 5
T-tropic HIV-infected T cell contact with AMs leads to HIV Gag staining in the AM cytosol. (A) AMs were co-cultured with HIV JR-CSF-infected autologous CD4+ T cells (cell-to-cell, CTC) or T cell supernatant (SN) for 3 h, washed, and cultured for an additional 14 days. Staining was performed with DAPI for cell nuclei, HIV Gag, CD3 for T cells, and CD68 for macrophages, and images were acquired by immunofluorescent confocal microscopy. White dotted outlines show the AM or T cell membrane. (B–D) % Gag+ cells out of all nucleated cells were quantified. (B) Gag + CD4+ T cells not associated with AMs (Gag+ T cells) were rare in both the SN and CTC conditions. (C) Cytosolic gag staining was increased in AMs associated with CD3+ T cells in the CTC condition compared to the SN condition. (D) Pie charts of all of the staining categories in SN and CTC conditions. Gag staining was rarely observed in solitary CD3+ T cells. Each category was compared between SN and CTC by the Friedman test with Dunn’s multiple comparison’s test. 100 cells per donor were quantified; n = 6 donors. *p < 0.05.
