FIG 5.

B cell-mediated trans infection of T_N is inhibited by anti-DC-SIGN. (A) B cells were incubated with 20 μg/ml anti-DC-SIGN MAb for 30 min at 4°C prior to pulsing with HIV-1^BaL (10⁻³ MOI) and then cocultured with CCL-19-treated T_N for trans infection as described in Materials and Methods. Supernatants were collected after 12 days and tested for HIV-1 Gag p24 by ELISA. B cells treated with mouse IgG (20 μg/ml) were used as the control. Data are mean ± SE, n = 3 replicates from 2 donors. (B) In parallel cultures, CCL-19-treated T_N were subsequently treated with maraviroc (1 μM) as described in Materials and Methods and cocultured with HIV-1^BaL (10⁻³ MOI)-pulsed B cells for trans infection. Supernatants were collected after 12 days and tested for HIV-1 Gag p24 by ELISA. Untreated T_N were used in the control cocultures. Data are mean ± SE, n = 3 replicates from 2 donors. Data were analyzed by one-way ANOVA followed by ad hoc Student’s t test. **, P < 0.005; ns, not significant. (C) Anti-DC-SIGN- or maraviroc-treated cells and the corresponding controls from the cocultures in panels A and B, as well as uninfected cocultures, were sampled after 12 days, stained with anti-Kc57, -CD4, -CD3, or -CD19, and analyzed by FACS as described in Materials and Methods.