Inhibition of HIV-1 replication in T lymphocytes by CNI-1493
(A) Immunoblot analysis of RAW 264.7 cells pretreated with varying concentrations of CNI-1493 1 hr prior to addition of LPS. Cell lysates were prepared for immunoprecipitation and immunoblotting (as described in Materials and Methods) 15 min after administration of LPS. (B–D) PHA-activated T lymphocytes were pretreated with control diluent or varying concentrations of CNI-1493 for 1 hr, infected with HIV-1LAI as in Fig. 1, and cultured for 12 days. Cells were refed every 3 days with fresh media containing IL-2 ± CNI-1493. Experiments were set up in duplicate to follow virus replication by measuring RT activity in the culture supernatants and to collect cell lysates at various time points for phospho-p38 MAPK immunoblotting. (B) Dose-dependent inhibition of RT by the addition of CNI-1493 on Day 6 of culture. (C) Time course of HIV-1 inhibition by 1 µM CNI-1493. (D) Immunoblot of phospho-p38 MAPK expression in HIV-1-infected cells on Day 6 after infection and treatment with 1 /µM CNI-1493.