Pattullo A, Patenaude P, Merzouki A, Sy T, Conway B, Montaner J, O Shaughnessy M, Cassol S; National Conference on Human Retroviruses and Related Infections.
Program Abstr Second Natl Conf Hum Retrovir Relat Infect Natl Conf Hum Retrovir Relat Infect 2nd 1995 Wash DC. 1995 Jan 29-Feb 2; 117.
B.C. Centre for Excellence in HIV/AIDS, St. Paul's Hospital, Vancouver, Canada.
Objective: To assess quantitative RT-PCR of HIV-1 spliced and unspliced mRNA using competitive RT-PCR and end-point dilution and plasma RNA by a commercial RT-PCR kit and to determine their utility in assessing therapy. Methods: Subjects with CD4 less than or equal to 400 enrolled in a randomized trial of combination AZT plus ddI had PBMC's collected at baseline and then monthly. RNA was extracted organically, DNase treated and reverse transcribed. Unspliced mRNA was quantified by end- point dilution PCR and by competitive PCR for a pol target using an engineered internal competitive template and an automated scanner. Results were compared with independent measures of plasma viremia as determined by the RocheR Amplicor HIV-1 Monitor RT PCR kit. Results: Full length (pol) mRNA was quantifiable in all patients at all time points with variability between subjects of several logs. Spliced regulatory mRNA was less abundant in most subjects and undetected in some. Plasma RNA was detected in nearly all samples with a range of 6 logs. There was high correlation between cellular pol mRNA, and plasma RNA copy numbers (R2=0.809). Significant changes in the levels of pol mRNA expression and plasma viremia were seen following imitation of antiretroviral therapy. The pattern of change seen by pol assay was mirrored by the plasma RNA assay. A more marked and persistent decline in HIV RNA occurred in naive patients while those previously treated had more transient and in some cases no decrease at all. Conclusion: There is high concordance in amount of cellular unspliced HIV-1 nRNA and plasma RNA by these assays confirming a direct relationship between virus expression and plasma virus load. Naive patients have a more marked and sustained response to treatment than those previously treated. These assays may offer a significant advance in the assessment of HIV disease activity and therapy.
Publication Types:
Keywords:
- Anti-HIV Agents
- Didanosine
- Drug Therapy, Combination
- Genes, pol
- HIV
- HIV Core Protein p24
- HIV Infections
- HIV Protease Inhibitors
- HIV Seropositivity
- HIV-1
- Humans
- Plasma
- Polymerase Chain Reaction
- RNA
- RNA, Viral
- Reverse Transcriptase Polymerase Chain Reaction
- Viremia
- Zidovudine
- drug therapy
- genetics
- immunology
- therapy
Other ID:
UI: 102213315
From Meeting Abstracts