To understand the persistence of latently HIV-1 infected cells in virally suppressed infected patients, a number of in vitro models of HIV latency have been developed. In an attempt to mimic the in vivo situation as closely as possible, several models use primary cells and replication-competent viruses in combination with antiretroviral compounds to prevent ongoing replication. Latency is subsequently measured by HIV RNA and/or protein production after cellular activation. To discriminate between pre- and post-integration latency, integrase inhibitors are routinely used, preventing novel integrations upon cellular activation. Here, we show that this choice of antiretrovirals may still cause a bias of pre-integration latency in these models, as unintegrated HIV DNA can form and directly contribute to the levels of HIV RNA and protein production. We further show that the addition of reverse transcriptase inhibitors effectively suppresses the levels of episomal HIV DNA (as measured by 2-LTR circles) and decreases the levels of HIV transcription. Consequently, we show that latency levels described in models that only use integrase inhibitors may be overestimated. The inclusion of additional control conditions, such as 2-LTR quantification and the addition of reverse transcriptase inhibitors, is crucial to fully elucidate the actual levels of post-integration latency.
- MeSH
- aktivace lymfocytů MeSH
- benzoxaziny farmakologie MeSH
- biologické modely MeSH
- CD4-pozitivní T-lymfocyty imunologie virologie MeSH
- DNA virů antagonisté a inhibitory biosyntéza genetika MeSH
- HIV infekce farmakoterapie imunologie virologie MeSH
- HIV-1 účinky léků fyziologie MeSH
- inhibitory HIV-integrasy farmakologie MeSH
- inhibitory HIV-proteasy farmakologie MeSH
- inhibitory reverzní transkriptasy farmakologie MeSH
- integrace viru účinky léků MeSH
- latence viru účinky léků MeSH
- lidé MeSH
- nevirapin farmakologie MeSH
- primární buněčná kultura MeSH
- raltegravirum kalicum farmakologie MeSH
- replikace viru účinky léků MeSH
- reportérové geny MeSH
- ritonavir farmakologie MeSH
- virové proteiny antagonisté a inhibitory biosyntéza genetika MeSH
- zelené fluorescenční proteiny genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- B-lymfocyty imunologie klasifikace MeSH
- buněčné linie MeSH
- DNA virů biosyntéza krev MeSH
- Gammaherpesvirinae genetika imunologie izolace a purifikace MeSH
- Herpesviridae imunologie MeSH
- imunofenotypizace MeSH
- myši MeSH
- slezina cytologie imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- MeSH
- DNA virů biosyntéza MeSH
- kuřecí embryo MeSH
- replikace viru MeSH
- virus vakcinie MeSH
- Check Tag
- kuřecí embryo MeSH
- MeSH
- DNA virů biosyntéza MeSH
- kuřecí embryo MeSH
- replikace viru MeSH
- virus vakcinie MeSH
- Check Tag
- kuřecí embryo MeSH
- MeSH
- DNA virů biosyntéza genetika MeSH
- mapování chromozomů MeSH
- molekulová hmotnost MeSH
- replikace DNA MeSH
- replikace viru * MeSH
- Retroviridae genetika metabolismus MeSH
- RNA transferová metabolismus MeSH
- RNA virová genetika MeSH
- virové geny * MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
