Fig. 3

GRFT in vivo release and activity. a For PK evaluation, GRT/CG FDIs were inserted vaginally in macaques either treated or not 4 weeks prior with DMPA. GRFT concentrations are shown in CVL (black symbols) and plasma (aqua symbols) at 4 h (left) and 8 h (right) post-insertion. Separate groups of macaques were assayed at each time point. The mean ± SEM is indicated by line and error bars for each group of 6 macaques. GRFT was not detected in plasma (PL) above the lower limit of quantification (LLOQ) of the assay, which was 10 ng/ml (top red dotted line), and so the values are shown at the LLOQ. The LLOQ for CVL was 1.25 ng/ml (bottom red dotted line). The 100-fold EC₉₀ level, 724.4 ng/ml, is also indicated (blue dotted line). Concentrations of GRFT in CVL from DMPA-treated and non-DMPA-treated macaques and between 4 and 8 h in DMPA-treated macaques were compared by two-sided Mann–Whitney test and p values are shown for α < 0.05. b GRFT concentrations in CVLs from non-DMPA-treated macaques and (c) from DMPA-treated macaques correlated with the EC₅₀ of the CVLs using Spearman correlation analysis. Spearman correlation coefficient (r) and significance of the association (p value) are shown. d Anti-SHIV SF162P3 activity of CVLs from non-DMPA-treated macaques was analyzed in human ectocervical explants. Tissue infection level (CUM SIV gag copies/ml) was compared between the Baseline (BL) and 4 h (4 h) post insertion using a log-normal mixed effects two-sided ANOVA model with time points and animal IDs nested within the experiment assumed as fixed and random effects, respectively. Four CVLs selected at random from the 6 macaques per time point were each tested twice for 8 replicates total. CVLs were collected from the same macaques at baseline and 4 h post-insertion. Mean ± SEM is indicated for each group by line and error bars