Fig. 5. No enrichment of HIV DNA in sorted CD32⁺ CD4⁺ T cells compared to CD32⁻ CD4⁺ T cells and CD32⁺ resting CD4⁺ T cells contribute minimally to the total pool of HIV DNA in CD4⁺ T cells.

(A) Measurements of HIV DNA were performed on CD32 pull-downs with antibody-conjugated magnetic beads and compared to cell number normalized pre-immunoprecipitation (pre-IP) pull-downs (n = 7) on fresh cells. (B) HIV DNA load was measured from CD32⁺ resting CD4⁺ T cells and CD32⁻ resting CD4⁺ T cells isolated using magnetic beads (n = 6). (C) FACS was performed to isolate total CD4⁺ T cells, CD32⁺ CD4⁺ T cells, CD32⁻ CD4⁺ T cells, CD32⁻ resting CD4⁺ T cells (HLA-DR⁻ CD69⁻ CD25⁻), CD32⁺ resting CD4⁺ T cells, CD32⁻ activated CD4⁺ T cells (HLA-DR⁺ or CD69⁺ or CD25⁺), and CD32⁺ activated CD4⁺ T cells from freshly isolated PBMCs from HIV⁺ ART-suppressed individuals, and HIV DNA load was measured in all sorted populations (n = 10). Each patient is represented by a different symbol. Lines represent median. (D and E) Contribution of each cell population to the total pool of HIV DNA in CD4⁺ T cells calculated in 10 HIV⁺ ART-suppressed individuals. HIV total DNA copy number was determined in sorted subsets by qPCR. Each symbol represents a different individual. The contribution of each subset to the total pool of HIV DNA in CD4⁺ T cells was calculated by accounting for the frequency of these subsets within the CD4⁺ compartment and abundance of HIV DNA determined in each subset. Horizontal lines indicate median values. All statistical comparisons were performed using two-tailed Wilcoxon rank tests. *P < 0.05 and **P < 0.01.