FIG. 4.

Rapamycin inhibits virus transcription in WE17/10 and primary human cells in an FK506-dependent manner. (A) WE17/10 cells (10⁵) were either left untreated or treated for 30 min with rapamycin (RAPA, 1.1 nM), FK506 (100 ng/ml), or both rapamycin and FK506 simultaneously. Cells were then either left uninfected or infected with HXB-Luc particles (10 ng of p24). (B) Freshly isolated PBLs fromthree healthy donors were kept in culture for 48 h in the presence of phytohemagglutinin protein (3 μg/ml) and rhIL-2 (50 U/ml). PBLs (10⁵) were then either left untreated or treated for 30 min with rapamycin (1.1 nM), FK506 (100 ng/ml), or both rapamycin and FK506 simultaneously. Cells were then either left uninfected or infected with HXB-Luc particles (10 ng of p24) or Ada-M-pseudotyped viruses. Cells were incubated at 37°C for 72 h and then lysed for monitoring luciferase activity. Data from only one representative donor are presented. Results shown are the means ± standard deviation of quadruplicate samples and are representative of at least three independent experiments.