Quantification and comparison of HIV-1 proviral load in peripheral blood mononuclear cells and isolated CD4+ T cells.
HIV proviral load was determined by quantitative DNA polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC) and lymphocyte subsets isolated by cell sorter. Provirus measured in PBMC, when expressed as HIV copy number per million CD4+ cells, resulted in values which approximated those obtained from sorted CD4+ T lymphocytes. A cross sectional analysis of HIV proviral load in CD4+ T cells from 25 previously untreated and 30 zidovudine-treated seropositive patients with CD4+ T-cell counts between 25 and 802/mm3 demonstrated HIV copy numbers ranging from 1 copy per 10,000 cells in early disease to 1 copy per 10 cells in advanced disease. HIV proviral load can be rapidly assayed by PCR to give a reproducible value which varies over a 1,000-fold range and is positively correlated with cell infectivity as measured by a quantitative micrococulture assay. A less technically demanding assay using PBMC as substrate can give similar results to those obtained with sorted CD4+ T cells.
Wood
R
R
Division of Infectious Diseases, Stanford University Medical Center, CA 94305.
Dong
H
H
Katzenstein
D A
DA
Merigan
T C
TC
D002452:Cell Count; D004279:DNA, Viral / Q000302:isolation & purification*; D006678:HIV / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000302:isolation & purification*; D013601:T-Lymphocytes / Q000166:cytology / Q000382:microbiology*
D004279:DNA, Viral