Dailey P, Wilber J, Neuwald P; International Conference on AIDS.
Int Conf AIDS. 1994 Aug 7-12; 10: 76 (abstract no. 580A).
Chiron Corporation, Emeryville, CA 94608.
OBJECTIVE: To evaluate the feasibility of quantitating HIV RNA in Peripheral Blood Mononuclear Cells (PBMCs), and examine the relationship between HIV viral load in these cells with viral load in plasma using a bDNA signal amplification assay. METHODS: Plasma and cell samples were obtained from 30 HIV seropositive patients attending an AIDS outpatient clinic. PBMCs were purified by Leukoprep density gradient separation and frozen at -80 degrees C as a dry pellet. Samples were homogenized in 8M guanidine HCl and 0.5% sarcosyl, and RNA was selectively precipitated with ethanol. The precipitate was resolubilized in the bDNA assay sample buffer containing Proteinase K and detergent and added directly to a 96-well microplate for quantitation of HIV RNA (Quantiplex HIV-RNA, Chiron Corporation) in the same manner as plasma samples. HIV RNA levels were expressed as HIV RNA Eq/ml in plasma and HIV RNA Eq/10(7) cells in PBMCs. RESULTS: HIV RNA was detected and quantitated in 89% and 83% of 30 paired plasma and PBMC specimens, respectively. There was a significant correlation between the plasma and PBMC HIV RNA levels (r = .54, p = .003). The mean ratio of plasma/PBMC HIV RNA levels for all patients was 13.1 (range: 0.1-115.0). This ratio increased with progression of disease. The plasma/PBMC ratio increased from 6.1 to 14.02 and then to 23.8 in samples from patients in CDC clinical categories A (asymptomatic), B (symptomatic), and C (AIDS), respectively. DISCUSSION AND CONCLUSIONS: The bDNA assay is a simple and convenient method which can be used to quantitate HIV viral load in PBMCs as well as plasma after a simple processing step. A significant correlation was observed between plasma and PBMC viral load. The plasma/PBMC HIV RNA ratio increased with disease progression. Evaluation of HIV viral load in both compartments of the peripheral blood may prove useful in evaluating antiviral therapy and studying the natural history and clinical progression of AIDS.
Publication Types:
Keywords:
- AIDS Vaccines
- Acquired Immunodeficiency Syndrome
- Anti-HIV Agents
- Branched DNA Signal Amplification Assay
- DNA
- Disease Progression
- Evaluation Studies
- HIV
- HIV Core Protein p24
- HIV Infections
- HIV Seropositivity
- Humans
- RNA
- RNA, Viral
- Viral Load
- immunology
- organization & administration
Other ID:
UI: 102211311
From Meeting Abstracts