Fig. 5: PRR851 impedes the productive infection of native Env-pseudotyped HIV-1 in activated CD4+ T cells.

a, b PHA/IL-2-activated primary CD4⁺ T cells were 30-min pretreated with 10 µM ICZ, PRR851, PRR846, or DMSO before RetroNectin-based, 6-h HIV-89.6-EGFP infection. Cells were washed and incubated for 48 h in the presence of drugs. Samples were processed for FC and histograms are displayed (a). DMSO-treated, noninfected cells were used for the gating. The percentage of EGFP⁺ cells is plotted (b n = 3). c, d Representation of alternative protocols (c) and quantification of the EGFP expression in infected PHA/IL-2-activated CD4⁺ T cells by FC (d n = 3 per condition). In addition to negative and positive controls, i.e., without (#i) or in constant presence of PRR851 (10 µM) (#ii), respectively, the drug was removed after 6-h infection (#iii) or added after (#iv). e, f PHA/IL-2-activated CD4⁺ T cells were 30-min pretreated with DMSO or 10 µM PRR851 before RetroNectin-based, 6-h HIV-89.6-EGFP infection, washed and further incubated for 48, 72 or 96 h in the presence of drugs. Samples were processed for FC for the expression of EGFP and HIV-1 Gag. Representative FC histograms are displayed (e). DMSO-treated, noninfected cells were used for the gating. The percentage of EGFP⁺, Gag⁺, or double-positive cells is plotted (f n = 3). Note that single EGFP⁺ cells (green boxes) decrease during incubation time, while double-positive cells increase in a drug-dependent manner (red boxes). g PHA/IL-2-activated CD4⁺ T cells were 30-min pretreated with DMSO or 10 µM PRR851 before 6-h HIV-89.6-EGFP infection by spinoculation instead of the RetroNectin method. After 48 h, samples were processed for FC and the percentage of EGFP⁺ cells is plotted (n = 3). We used a MOI of 2. In all cases, means ± S.D. and individual value are presented. P values are indicated.