(A–N) LN resident and migratory DCs were isolated from treated aviremic HIV-infected individuals.
(A) Frequencies of LN DCs harboring integrated HIV DNA (cells per million) directly ex vivo (N = 4).
(B) Frequencies of LN DCs harboring cell-associated unspliced HIV gag RNA or multi-spliced tat-rev HIV RNA (cells per million) directly ex vivo (N = 3).
(C) Levels of HIV RNA (copies/mL) produced in culture supernatants of unstimulated VOA conditions (N = 3).
(D) Levels of HIV RNA (copies/mL) produced in culture supernatants of stimulated VOA conditions (N = 9).
(E) Proportion of HIV-infected individuals with detectable HIV RNA (≥20 HIV RNA copies/mL) in stimulated VOA conditions at day 14 (N = 9).
(F) Levels of HIV RNA (copies/mL) produced in the culture supernatants at day 14 in the stimulated VOA conditions (N = 9).
(G) Correlation between duration of ART (in years) and levels of HIV RNA (copies/mL) produced in the stimulated VOA culture supernatants at day 14 (N = 9).
(H–J) (H) Estimated decay slopes for HIV-RNA levels in stimulated VOA culture supernatants of LN CD4 T cells (H), LN migratory DCs (I), or LN resident DCs (J) with the duration of suppressive therapy.
(K) Proportion of HIV-infected individuals with detectable HIV-1 RNA (≥20 HIV RNA copies/mL) in LN cell populations at day 14 of the in vitro HIV-1 infection assay (N = 9).
(L) Levels of HIV RNA (copies/mL) in culture supernatants at day 14 in the in vitro infection assay (N = 9).
(M) Schematic representation of the QVOA preformed on LN resident and migratory DCs. HIV-infected individuals were considered as replicates at 40,000 cell concentration (N = 9).
(N) Estimated mean frequencies of LN resident and migratory DCs harboring inducible replication-competent virus/RNA units per million (RUPM) cells.
Dashed line represents the limit of detection (A–D), (F), and (L). Undetectable values were arbitrarily defined as 10 HIV RNA copies/mL (C), (D), (F–J), and (L). HIV-infected individuals were color coded (A), (B), (F), and (L). Histograms correspond to the estimated mean (E), (F), (K), (L), and (N). Red bars correspond to mean ± SEM (D), (F), and (L) or to lower and upper confidence interval at 0.95 (N). Red stars indicate statistical significance (*p < 0.05) (D–F) and (K). Statistical significance (p values) was obtained using one-way ANOVA (Kruskal-Wallis test) followed by Wilcoxon matched-pairs one-tailed signed rank test (D) and (F) or using Spearman rank test for correlations (G) or using chi-square test for proportion of positive individuals (E) and (K). ‘‘Estimate’’ refers to the decay slopes for LN CD4 T cells (H), LN migratory DCs (I), or LN resident DCs (J). Red color wells correspond to HIV-RNA-negative wells, whereas green wells correspond to HIV-RNA-positive wells (M). ‘‘Res DC’’ corresponds to LN resident DCs and ‘‘Mig DC’’ corresponds to LN migratory DCs.