Fig. 3

Ablation of CCR5 usage reduces dual-tropic HIV-1 pathogenesis in PBMC infection. a Intracellular p24 staining and FACS analysis was used to quantify viral replication in infected PBMCs. Infected (p24+) CD4 T cell numbers were calculated by multiplying the percentage of (p24+) cells with total CD4 T cell numbers at each time point (left). RT-qPCR analysis of HIV-1 genomic RNA levels was used to measure extracellular viral replication of R3A and R3A-5/6AA in infected PBMCs (right). b Representative FACS Plots are presented showing CD4 and CD8 populations gated on live CD3(+) T cells in PBMC at 6 days post infection. HIV-1 fusion inhibitor T20 treatment was used as negative control. c Kinetics of CD4 T cell depletion by R3A versus R3A-5/6AA was analyzed as described in Fig. 1b. CD4 T cell survival are measured as %CD4 T cells in infected PBMCs relative to mock infection (top), or live CD4 T cell numbers (bottom) over time. d p24(−) and p24(+) CD4 T cell death in R3A and R3A-5/6AA infected PBMCs are quantified at 6 days post infection, as described in Fig. 1C. FACS plots (left) and graphical summary (Right) from a representative experiment are presented. Experimental results were repeated with PBMCs from 3 different blood donors. *p < 0.05