In vivo evidence for instability of episomal human immunodeficiency virus type 1 cDNA.
Current regimens for the management of human immunodeficiency virus type 1 (HIV-1) infection suppress plasma viremia to below detectable levels for prolonged intervals. Nevertheless, there is a rapid resumption in plasma viremia if therapy is interrupted. Attempts to characterize the extent of viral replication under conditions of potent suppression and undetectable plasma viremia have been hampered by a lack of convenient assays that can distinguish latent from ongoing viral replication. Using episomal viral cDNA as a surrogate for ongoing replication, we previously presented evidence that viral replication persists in the majority of infected individuals with a sustained aviremic status. The labile nature of viral episomes and hence their validity as surrogate markers of ongoing replication in individuals with long-term-suppressed HIV-1 infection have been analyzed in short-term in vitro experiments with conflicting results. Since these in vitro experiments do not shed light on the long-term in vivo dynamics of episomal cDNA or recapitulate the natural targets of infection in vivo, we have analyzed the dynamics of episomal cDNA turnover in vivo by following the emergence of an M184V polymorphism in plasma viral RNA, in episomal cDNA, and in proviral DNA in patients on suboptimal therapies. We demonstrate that during acquisition of drug resistance, wild-type episomal cDNAs are replaced by M184V-harboring episomes. Importantly, a complete replacement of wild-type episomes with M184V-containing episomes occurred while proviruses remained wild type. This indicates that episomal cDNAs are turned over by degradation rather than through death or tissue redistribution of the infected cell itself. Therefore, evolution of episomal viral cDNAs is a valid surrogate of ongoing viral replication in HIV-1-infected individuals.
Sharkey
Mark
M
Program in Molecular Medicine, University of Massachusetts, 373 Plantation St., Biotech 2, Suite 319, Worcester, MA 01605, USA.
Triques
Karine
K
Kuritzkes
Daniel R
DR
Stevenson
Mario
M
D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000188:drug therapy; D019380:Anti-HIV Agents / Q000627:therapeutic use; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D042822:Genomic Instability / Q000235:genetics*; D015658:HIV Infections / Q000097:blood / Q000188:drug therapy; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D012367:RNA, Viral / Q000235:genetics; D018894:Reverse Transcriptase Inhibitors / Q000627:therapeutic use; D014766:Viremia / Q000097:blood
D019380:Anti-HIV Agents; D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral; D018894:Reverse Transcriptase Inhibitors