Fig. 5

Preferential loss of resting CD4 T cells infected with integrated proviruses. a Productively infected GFP+ cells containing Int-WT or Int-D116N HIV-1 were sorted by FACS 7 days after infection and placed back into culture. Survival of the GFP+ cells was assessed by flow cytometry on the indicated days, and virus production from the FACS purified GFP+ cells was measured by RT-qPCR for viral RNA present in the culture medium as described [6]. In this experiment an Envelope+ virus was employed. b Experimental design for c–e. Resting CD4 T cells infected with HSA-reporter viruses were maintained in IL-4 ± RAL for 14 days. At the indicated time points HSA⁺ cells in each sample were isolated using anti-HSA antibody coupled to magnetic beads. Efavirenz was added to all cells on day 8. HSA Positive and negative cells were separated using the MACS system. Cell survival was measured by forward and side scatter analysis. Data present one of two independent experiments. c Productively infected HSA⁺ cells purified from No RAL cultures at day 5, 7 and 14 p.i. were analyzed for viral DNA. d The ratios of 2-LTR/total HIV-1 DNA in comparison with the ratio of integrated/total HIV-1 DNA for No RAL purified HSA⁺ productively infected cells. e HSA⁺ and HSA-neg. cells were purified at day 7 and 14 p.i. and placed into fresh medium. Cell viability of these individual populations was monitored by flow cytometry over 48 h