FIG 5.

HIV replication is inhibited before translation in M1- and M2-polarized macrophages and in TLR-primed macrophages. (A) Polarized and TLR-primed macrophages were inoculated by spinoculation (centrifugation at 1,200 × g for 2 h at room temperature) with HIV env-pseudotyped reporter (luciferase) viruses and 24 h later were analyzed for luciferase activity (n = 3). ADA is a well-described envelope, and env #8 is the envelope of a primary isolate cloned by our laboratory. Both were obtained using CCR5. (B) 3M-002 (TLR8 agonist) was added to the culture medium either for 24 h before or immediately after HIV challenge with VSV env-pseudotyped reporter (luciferase) (the same inoculation protocol used for HIV env-pseudotyped viruses). After 24 h, the MDMs were analyzed for luciferase activity. n.s., not significant. (C) Analysis of integrated HIV DNA by Alu PCR with primers specific for human Alu sequences and for HIV-1-based lentiviral vector sequences. MDMs were pretreated with the various TLR agonists and infected with the replication-competent HIV strain YU-2, and DNA was extracted 24 h later. The agonists are described in the legend to Fig. 1.