Figure 1
MPA increases R5-tropic HIV-1 replication in cervical explants in a dose-dependent manner and at peak serum levels of DMPA-IM users. (a–c) Ectocervical or (d,e) endocervical explants for each donor were treated in parallel with vehicle (EtOH [0.1% v/v]) or 1, 10 or 100 nM MPA for 48 hr after which tissue was infected. washed and left for varying times in the presence of vehicle or the different concentrations of MPA. Media were collected at 3, 5, 7 and 10 days post infection and assayed for p24 levels by ELISA. Infection with R5 HIV-1BaL-Renilla was measured and relative infection was plotted relative to day 3 vehicle set to 1. Relative infection for pooled data was used to compare the fold-change in infection for matched incubations with vehicle or MPA, since the absolute infection levels as measured by p24 assays at day 3 varied in a donor-dependent manner. Each condition was performed at least in triplicate. For data set (a), seven of these were performed as independent donor experiments comparing only MPA vs vehicle in parallel. An additional 10 experiments were performed independently with MPA vs NET vs vehicle, used in Fig. 2a, and the MPA vs vehicle data only from these 10 data sets were also used in (a). MPA vs vehicle data from three experiments shown in Fig. 3b, were also used in (a), making the total for n = 20 independent donor data sets shown in (a). Results of b and c are representative of (b) nine and (c) five independent donor experiments. (d) MPA vs vehicle data are from six experiments done in parallel with NET, also shown in Fig. 2b (e). Results are shown from 5 independent donor experiments. XY plots are representative of mean + SEM. Statistical significance was determined by comparing MPA to the vehicle control using a repeated measures two-way ANOVA with a post hoc Sidak’s test, with *, **, *** and **** denoting p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively. Detailed statistical analysis is shown in Supplementary Tables S3 and S4. All data sets except one had all data time points. For (a) one of the donors (49) experiments was missing a day 5 value and was therefore excluded from the statistical analysis, although it is included in the final Figure.
