期刊
CELL REPORTS METHODS
卷 2, 期 6, 页码 -出版社
CELL PRESS
DOI: 10.1016/j.crmeth.2022.100238
关键词
-
资金
- NIH/NIAID [R01AI136916]
- NIH [AI150355]
There is no cure for HIV infection due to the establishment of a latent reservoir that escapes antiretroviral treatments. A viral vector has been developed to accurately identify cells with latent proviruses and reveal mechanisms of latency establishment.
There is no cure for HIV infection, as the virus establishes a latent reservoir, which escapes highly active antiretroviral treatments. One major obstacle is the difficulty identifying cells that harbor latent proviruses. We devised a single-round viral vector that carries a series of versatile reporter molecules that are expressed in an LTR-dependent or LTR-independent manner and make it possible to accurately distinguish productive from latent infection. Using primary human CD4(+) T cells, we show that transcriptionally silent proviruses are found in more than 50% of infected cells. The latently infected cells harbor proviruses but lack evidence for multiple spliced transcripts. LTR-silent integrations occurred to variable degrees in all CD4(+) T subsets examined, with CD4(+) TEM and CD4(+) TREG displaying the highest frequency of latent infections. This viral vector permits the interrogation of HIV latency at single-cell resolution, revealing mechanisms of latency establishment and allowing the characterization of effective latency-reversing agents.
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