New Assay Reveals Vast Excess of Defective over Intact HIV-1 Transcripts in Antiretroviral Therapy-Suppressed Individuals
| Autor: | Holly Anne Martin, Gayatri Nikhila Kadiyala, Sushama Telwatte, Adam Wedrychowski, Tsui-Hua Chen, Sara Moron-Lopez, Doug Arneson, Rebecca Hoh, Steven Deeks, Joseph Wong, Steven A. Yukl |
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| Přispěvatelé: | Kirchhoff, Frank |
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
HIV RNA
latent infection intact Immunology HIV Infections Medical and Health Sciences Microbiology Proviruses HIV DNA Virology Genetics Humans transcriptional regulation Viral HIV provirus defective Agricultural and Veterinary Sciences Biological Sciences Genome Replication and Regulation of Viral Gene Expression infectious Infectious Diseases Good Health and Well Being intact proviral DNA assay Insect Science HIV-1 RNA viral replication HIV/AIDS transcription Infection Biotechnology |
| Zdroj: | J Virol Journal of virology, vol 96, iss 24 |
| Popis: | Most of the HIV DNA in infected individuals is noninfectious because of deleterious mutations. However, it is unclear how much of the transcribed HIV RNA is potentially infectious or defective. To address this question, we developed and validated a novel intact viral RNA assay (IVRA) that uses droplet digital reverse transcriptase PCR (dd-RT-PCR) for the commonly mutated packaging signal (Psi) and Rev response element (RRE) regions (from the intact proviral DNA assay [IPDA]) to quantify likely intact (Psi(+) RRE(+)), 3′ defective (Psi(+) RRE(−)), and 5′ defective (Psi(−) RRE(+)) HIV RNA. We then applied the IPDA and IVRA to quantify intact and defective HIV DNA and RNA from peripheral CD4(+) T cells from 9 antiretroviral therapy (ART)-suppressed individuals. Levels of 3′ defective HIV DNA were not significantly different from those of 5′ defective HIV DNA, and both were higher than intact HIV DNA. In contrast, 3′ defective HIV RNA (median 86 copies/10(6) cells; 94% of HIV RNA) was much more abundant than 5′ defective (2.1 copies/10(6) cells; 5.6%) or intact (0.6 copies/10(6) cells |
| Databáze: | OpenAIRE |
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