Figure 5. LN DCs of viremic individuals are HIV-infected, transcriptionally active, and produce HIV upon TLR7/8 in vitro stimulation.

(A–F) LN resident and migratory DCs were isolated from untreated viremic HIV-infected individuals.

(A) Levels of integrated HIV DNA (levels/million cells) within LN resident and migratory DCs (N = 5).

(B) Levels of cell-associated unspliced HIV gag RNA within LN resident and migratory DCs (N = 5).

(C and D) (C) Frequencies of cells containing unspliced HIV gag RNA (cells/million) (C) and multi-spliced tat-rev RNA (cells/million) (D) in LN resident and migratory DCs (N = 5).

(E and F) (E) Levels of HIV production in LN resident and migratory DC culture supernatants at day 7 post-stimulation with TLR7/8 agonist as assessed by HIV-RNA levels (copies/mL) (E) or HIV P24 levels (pg/mL) (F) (N = 6).

Autologous LN PD-1⁺/Tfh and LN PD-1⁻/non-Tfh cells were used as controls (A–F). LN PD-1⁺/Tfh and LN PD-1⁻/non-Tfh cells were stimulated or not with anti-CD3/anti-CD28 mAbs for 3 days (E and F). Dashed line represents the limit of detection (A–F). HIV-infected individuals are color coded (A–D). Red bars correspond to mean ± SEM (A–F). Red stars indicate statistical significance (*p < 0.05). Statistical significance (p values) was obtained using one-way ANOVA (Kruskal-Wallis test) followed by Wilcoxon matched-pairs one-tailed signed rank test (E and F). ‘‘Res DC’’ corresponds to LN resident DCs and ‘‘Mig DC’’ corresponds to LN migratory DCs.