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6 záznamů v Medvik Filtry

Článek

Protein expression from unintegrated HIV-1 DNA introduces bias in primary in vitro post-integration latency models

Bonczkowski, Pawel
Autor Bonczkowski, Pawel HIV Cure Research Center, Department of Internal Medicine, Faculty of Medicine and Health Sciences, Ghent University and Ghent University Hospital, Ghent, Belgium
De Scheerder, Marie-Angélique
Autor De Scheerder, Marie-Angélique HIV Cure Research Center, Department of Internal Medicine, Faculty of Medicine and Health Sciences, Ghent University and Ghent University Hospital, Ghent, Belgium
Malatinkova, Eva
Autor Malatinkova, Eva HIV Cure Research Center, Department of Internal Medicine, Faculty of Medicine and Health Sciences, Ghent University and Ghent University Hospital, Ghent, Belgium
Borch, Alexandra
Autor Borch, Alexandra Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, 63225 Langen, Germany
Melkova, Zora
Autor Melkova, Zora Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, 63225 Langen, Germany. Department of Immunology and Microbiology, 1st Medical Faculty, Charles University, Studnickova 7, 128 00 Prague 2, Czech Republic
Koenig, Renate
Autor Koenig, Renate Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, 63225 Langen, Germany. Immunity and Pathogenesis Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA
De Spiegelaere, Ward
Autor De Spiegelaere, Ward HIV Cure Research Center, Department of Internal Medicine, Faculty of Medicine and Health Sciences, Ghent University and Ghent University Hospital, Ghent, Belgium. Department of Morphology, Faculty of Veterinary Sciences, Ghent University, Merelbeke, Belgium
Vandekerckhove, Linos
Autor Vandekerckhove, Linos HIV Cure Research Center, Department of Internal Medicine, Faculty of Medicine and Health Sciences, Ghent University and Ghent University Hospital, Ghent, Belgium

Scientific reports. 2016 ; 6 (-) : 38329. [pub] 20161202

Sci Rep
ISSN 2045-2322
Medvik
Zdroj

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.