Figure 4.

CD4⁺ T cells from PLWH on ART that survive ex vivo venetoclax co-culture have reduced levels of total and intact HIV-1 pro-viral DNA and exhibit a less apoptotic transcriptional signature

Total CD4⁺ T cells isolated from peripheral blood of PLWH on ART were co-cultured with venetoclax (VNX) or DMSO control for 24 h before washing and harvesting 24 h later for quantification of HIV-1 DNA. For flow cytometry analysis and RNA sequencing (RNA-seq), cells were harvested immediately following 24 h VNX co-culture.

(A) Fold change in total integrated HIV-1 DNA per million CD4⁺ T cells relative to DMSO control (n = 6 donors).

(B) Absolute frequency of total integrated HIV-1 DNA per million CD4⁺ T cells for the 100 nM dose.

(C) Fold change in intact HIV-1 DNA per million CD4⁺ T cells (n = 11 donors).

(D) Absolute frequency of intact HIV-1 DNA per million CD4⁺ T cells for the 100 nM dose.

(E) Heatmap visualizing the normalized proportion of dead/dying Violet⁺ cells within stated CD4⁺ T cell subsets at different VNX concentrations (n = 5 donors).

(F and G) Normalized expression values by VNX treatment group for (F) pro-death and (G) pro-survival human Bcl-2 family genes. NA, naive; CM, central memory; EM, effector memory; TM, transitional memory; TD, terminally differentiated. For (B) and (D), MFC, median fold change.

p values for (A) and (C) were calculated using a one-sample Wilcoxon signed-rank test. p values for (B) and (D) were calculated using a Wilcoxon matched-pairs signed-rank test. Error bars in (A) and (C) show median ± 95% confidence interval (CI). Boxplots in (F) and (G) show median ± interquartile range. Each symbol represents a different donor.