(A) Bacterial culture supernatants (1:4 dilution) were applied to Jurkat cells for 1 hour prior to the addition of 0.5 μM [³H]TFV or [³H]TDF and Jurkat cell–associated drug quantified by liquid scintillation counting. (B) Supernatants from indicated strains of G. vaginalis, A. vaginae, L. crispatus, L. iners, or P. bivia were diluted 1:8 to 1:64 in PBS and applied to cells for 1 hour prior to addition of 50 μM [¹⁴C]TFV. Data are presented as mean ± SEM from at least 2 independent experiments with duplicate measurements per condition. (C) Different human cell lines or primary CD4⁺ T cells were exposed to 1:8 dilution of A. vaginae (DNF00720) or 1:32 dilution of G. vaginalis (DNF00162) supernatants for 1 hour prior to exposure to 0.5 μM radiolabeled drugs. Results are mean ± SEM of duplicate measurements from at least 2 independent experiments. (D) Bacterial culture supernatants (1:4 dilution) were applied to Jurkat cells for 1 hour prior to the addition of radiolabeled adefovir (ADF), adefovir dipivoxil (ADP), or dapivirine DPV and cell-associated drug quantified by scintillation counting. Results are mean ± SEM of duplicate measurements from 2 independent experiments. (E) Jurkat cells were incubated with the indicated culture supernatants, abiotic control broth, 80 μM dynasore, or PBS prior to exposure to 50 μg/ml of Alexa Fluor 555–conjugated transferrin (Tf-555) in the presence of the supernatants. The plasma membranes were stained green with DiO, nuclei blue with DAPI, and the transferrin is red. Representative images from 2 independent experiments are shown. (F) Approximately 100 cells from 6–9 randomly selected fields were counted and the percentage of cells that internalized Tf-555 is shown. The asterisks indicate significant differences relative to abiotic controls by unpaired Student’s t test (A and F) or by ANOVA with Tukey’s multiple-comparison correction (C and D). *P < 0.05; ***P < 0.001; ****P < 0.0001.