title: Expression of CD25 defines peripheral blood mononuclear cells with productive versus latent HIV infection.
abstract: The present studies were designed to further determine 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 CD25high and CD25low 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.
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235, USA.
authors.forename: J
authors.initials: J
authors.lastname: Borvak
authors.forename: C S
authors.initials: CS
authors.lastname: Chou
authors.forename: K
authors.initials: K
authors.lastname: Bell
authors.forename: G
authors.initials: G
authors.lastname: Van Dyke
authors.forename: H
authors.initials: H
authors.lastname: Zola
authors.forename: O
authors.initials: O
authors.lastname: Ramilo
authors.forename: E S
authors.initials: ES
authors.lastname: Vitetta
mesh_terms: D001483:Base Sequence; D015415:Biomarkers; D002478:Cells, Cultured; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D005455:Fluorescent Antibody Technique; D015658:HIV Infections / Q000276:immunology*; D006801:Humans; D000922:Immunotoxins; D007963:Leukocytes, Mononuclear / Q000276:immunology* / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015375:Receptors, Interleukin-2 / Q000032:analysis* / Q000276:immunology
chemical_list: D015415:Biomarkers; D017931:DNA Primers; D004279:DNA, Viral; D000922:Immunotoxins; D015375:Receptors, Interleukin-2