HCMV/HIV-1 coinfected PB-CD34+ cells and enhanced transfer of HIV-1 to autologous CD4+ T cells by latent HCMV. PB-CD34 cells were infected with mock or HCMV strain Merlin for 5 days before HIV-1NL4-3 infection for 3 days. (A) GM-CSF/TNF-α reactivation treatment for 14 hours to detect HIV-1 Gag p24 (red) and HCMV gB (green) by immunofluorescence and counterstained for nucleus (Hoechst, blue). Representative images with bright field and a scale bar of 10 μm are shown. (B) Flow cytometry detection of p24 signals analyzed on gB+- (shaded) and gB−-gated (dotted) for HCMV-infected samples. Representative histograms of 2 independent experiments are shown. (C) After 7 days post-HIV infection of mock or HCMV PB-CD34 cells, coculture with autologous naïve CD4+ T cells (1:5) was performed for 48 hours before being immunostained for flow cytometry. Representative plots with arrows indicating gating for analyzing intracellular p24 signals in CD4+ CD25+, or CD25− subpopulations in mock or HCMV- and/or HIV-1–infected PB-CD34 cells are shown. (C) Column graph for data from 4 independent experiments as a column graph showing mean ± SEM. Numbers represent percentages. *P < .05, **P < .01.
