Figure 5

Increase in TERT-2 cell-associated, infectious HIV-1 independent of direct interactions with P. gingivalis and blocked by anti-CCR5. (A) P. gingivalis was co-cultured with R5-HIV-1 on glass slides. (B and C) TERT-2 cells were pre-incubated with P. gingivalis, washed and inoculated with R5-HIV (Ba-L), washed, fixed and permeabilized for confocal microscopy analysis. Cells were stained with antibodies (1:100 dilutions) against P. gingivalis and HIV p24, or isotype control IgG. The confocal analysis was validated for assessment of intracellular HIV-1 using TZM-bl cells. Color key: Blue, DAPI; Red, Alexa 568; and Green, FITC conjugated IgG. Scale bars: 5 μm (A), 10 μm (B and C). Pictures are representative of 2 independent experiments and show one z-slice through the middle of the nucleus. (D), TERT-2 cells were pre-incubated with P. gingivalis or untreated and then incubated with anti-CCR5 antibody for 1 h. All TERT-2 cell cultures were then inoculated with R5-HIV-1. Culture supernatants (gray) and cell lysates (black) were collected at 18 hours post-inoculation with P. gingivalis as described in the legend of Fig. 1. Infectious virions were estimated using TZM-bl reporter cells. Data represent the mean ± SEM of 3 independent experiments, each performed in triplicate.