FIG. 3.
Depletion of cholesterol inhibits HIV-1 entry into CD4+ T cells independently of virus tropism. HIV-1 entry into Jurkat T cells (A and B) and PM1 T cells (C and D) was analyzed by semiquantitative PCR. Cells untreated, MβCD treated, or MβCD treated and reconstituted with cholesterol for 1 h at 37°C (MβCD/cholesterol) were infected (5 cpm of RT/cell) with X4 NL4-3 (A and C) or R5 Ad8 HIV-1 (D). One hour after infection, the cells were lysed, and PCR amplification of serially diluted cell lysates was performed with R/U5 primers specific for strong-stop DNA as described in Materials and Methods. Amplification of the β-globin gene was used to control the amount of DNA in each sample. PCR products were resolved by electrophoresis in 2% agarose gels and stained with ethidium bromide. (B) Inhibition of HIV-1 entry into Jurkat cells pretreated with monoclonal anti-CD4 antibody Q4120 (10 μg/ml) or with an antagonist of CXCR4 receptor, AMD3100 (1 μg/ml). (C) AMD3100 also inhibits HIV-1 NL4-3 entry into PM1 T cells.