FIG 2.

Treatment with intact Neisseria gonorrhoeae and Escherichia coli represses HIV-1 replication at the level of transcription. (A) MDMs were infected with a single-round, replication-defective HIV-luciferase reporter virus and, 48 h after infection, were cultured overnight with increasing amounts of gonococcus (GC). Cells were then lysed and assayed for luciferase activity. Bars represent mean (± SD) of seven donors; each donor was tested in triplicate. (B) MDMs were infected as in panel A and, 48 h after infection, were cultured with increasing amounts of live or heat-killed (56°C treatment) GC overnight. The cells were then lysed, and luciferase activity was measured. The data are the mean (± SD) of three donors; each donor was tested in triplicate. (C) MDMs were infected as described above and, 48 h after infection, were treated with heat-killed GC (HKGC) at a multiplicity of infection (MOI) of 10 for 24 h. Cells were then lysed and assayed for viral RNA accumulation by RT-qPCR. Shown are data from four donors. (D) MDMs were infected as in panel A and, 48 h after infection, cells were treated with heat-killed GC (MOI = 10) for 4 h. Cells were then treated with actinomycin D (10 μg/ml) to inhibit transcription. Total cytoplasmic RNA was prepared from the treated cultures at the indicated time points following actinomycin D treatment and analyzed by RT-qPCR for the expression of HIV-1 RNA. The data are the means (± SD) from four donors. (E) MDMs were infected as in panel A and, 48 h after infection, the cells were cultured overnight with increasing amounts of E. coli. Cells were then lysed and assayed for luciferase activity. Bars represent mean (± SD) of four donors, with each donor tested in triplicate. (F) MDMs were infected as in panel A and, 48 h after infection, cells were treated with heat-killed GC (MOI = 10) or heat-killed E. coli (HKEC) (MOI = 10) for 18 h. The cells were then lysed and assayed for luciferase activity. Bars represent the mean (± SD) of five male donors and five female donors; each donor was tested in triplicate. Although virus replication was decreased overall in MDMs from female donors compared to that in MDMs from male donors, it was repressed by HKGC and HKEC in a manner similar to that seen in MDMs from male donors. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.