Nef promotion of HIV-1 replication in primary resting CD4⁺ T cells is associated with T cell proliferation.

<p>Resting CD4⁺ T cells were isolated from PBMCs using immunomagnetic particles and were cultured in the presence of IL-2 before and during HIV-1 infection. (A) Resting CD4<b>⁺</b> T cells were cultured for 8 days and then infected with 5 ng p24 of replication-competent WT or Nef-mutated HIV-1. HIV-1 p24 production in the supernatants was measured at the times indicated. Representative data from one donor are shown. (B) Resting CD4<b>⁺</b> T cells were cultured for 8 days and then infected with 5 ng p24 of WT HIV-1 or Nef-mutated viruses. HIV-1 p24 production in the supernatant was measured at the times indicated. Mean values of two independent experiments on different donors are shown. (C) CFSE-labeled resting CD4<b>⁺</b> T cells (cultured for 8 days in the presence of IL-2) were mock infected or infected with 5 ng p24 of WT or Nef-mutated HIV-1. CD4<b>⁺</b> T cell proliferation was measured by flow cytometry at the indicated times. Data represent three independent experiments using cells from three different donors. Each symbol in the plots represents a single experiment result and the horizontal bars in the plots indicate the mean values of three independent experiments. There is no statistically significant difference in T-cell proliferation among WT and Nef-mutated HIV-1 infected CD4⁺ T cells.