Expression of CD25 defines peripheral blood mononuclear cells with productive versus latent HIV infection

The present studies were designed to further deter mine whether the CD25 marker could distinguish between cells productively and latently infected with HIV. This was accomplished by combining immunotoxin (IT)-mediated killing of CD25⁺ cells, highly sensitive indirect immunofluorescence to detect remaining CD25⁺ cells, and PCR-mediated amplification of proviral DNA in immunotoxin treated vs untreated HIV-infected cells. Our results demonstrate that: 1) By direct immunofluorescence 3 to 8% of PBMCs are CD25⁺, whereas by indirect immunofluorescence 30% are CD25⁺. The increased number of CD25⁺ cells is due to their detection by the highly sensitive indirect immunofluorescence assay. Up to 60% of the CD25⁺ cells are CD4⁺ and 12% are CD8⁺. 2) Treatment of HIV-infected PBMCs with an anti CD25 IT for 6 days eliminated both CD25(high) and CD25(low) cells and decreased the production of p24 by 99%. 3) Differences in the HIV proviral genome were detected in the unfractionated PBMCs vs PBMCs from which CD25⁺ cells had been eliminated by IT treatment. Hence, PBMCs containing both CD25⁺ and CD25^- cells express all intermediate proviral species and full-length double-stranded proviral DNA. In contrast, CD25^- quiescent cells contain predominantly intermediate species. These results confirm and extend our previous observations that expression of CD25 can distinguish latently infected cells from cells producing virus.