3rd Conference on Retroviruses and Opportunistic Infections Washington, DC - January 28-February 1, 1996

Conf Retroviruses Opportunistic Infect 1996 Jan 28-Feb 1; 3rd:61 (abstract no. 44)

Thomson MM¹, Vallejo A², Reitz MS¹, Biggar RJ¹, Gallo RC¹, Klotman ME³
NCI, NIH, Bethesda, MD.

Five HIV-1 infected individuals showing decline in CD4 cell counts after a prolonged period of stability were examined for expression of tat, rev and nef mRNAs in PBMCs, longitudinally collected before and after CD4 decline, by a reverse transcription-nested PCR assay. A fluorescent upper primer was used in the second PCR, and downstream primers were designed to result in products of different sizes depending on the splice acceptors used. An ABI 373A instrument was used for denaturing urea PAGE and analysis of amplified products, which were identified according to their length, determined relative to size standards run in the same lane. A total of 25 HIV-1 PCR products were detected in the 17 samples tested, 8 corresponding to nef, 12 to rev and 5 to tat. Nef was present in all of the samples, rev in 13 and tat in 12. The 4 rev- samples were all from time points prior to disease progression and were associated with a relatively high CD4 count. Singly spliced transcripts were undetectable in these samples by RT-PCR, while they were present in 11 of 13 rev+ samples. We conclude that, similar to what has been reported in vitro, a great variety of HIV-1 alternatively spliced transcripts are produced in vivo. Some patients in early stages of the infection appear to have undetectable expression of rev and of incompletely spliced mRNAs.

960128
44

Copyright © 1996 - Foundation for Retrovirology and Human Health . Reproduction of this abstract (other than one copy for personal reference) must be cleared through the Foundation for Retrovirology and Human Health.