FIG. 1.
Maturation dependence of HIV-1 infection in human MDMs. (A) HIV vector transduction. Adherent PBMCs were plated at 104 cells/well of a 96-well plate and infected with CMVΔR8.2(VSV-G) and CMVΔR8.9(VSV-G) packaged vectors (8.2-G and 8.9-G, respectively) or CMVΔR8.9(MLV-A) packaged vector [8.9(MLVA)] at an MOI of 10, as standardized on human 293T cells, on various days postisolation. Cells were cultured for 7 days posttransduction; percent cell transduction was then quantified by fluorescence-activated cell sorting. MLV vectors pseudotyped with VSV-G (MLV-G) were used as a control at the same MOI. Values are means ± standard errors of the means for three separate experiments. (B) HIV-1 infection. Titers of NL4-3 pseudotyped with VSV-G (NL4-G) were determined as for panel A, with the MOI standardized on 293T cells. Infection was scored by in situ p24 immunoassay. (C) Effect of cPPT on vector transduction (cPPT− or cPPT+). Titers of CMVΔR8.9(VSV-G) particles packaging pHR′-CMV-eGFP or HR′-cPPT-CMV-eGFP were determined as for panel A. The levels of p24 in both supernatants were equal (not shown), and infections were performed with an equivalent volume for which the cPPT− vector gave an MOI of 10 to standardize for virion input.