HIV latency in isolated patient CD4+ T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing

Steven A. Yukl, Philip Keiser, Peggy Kim, Sushama Telwatte, Sunil K. Joshi, Mai Vu, Harry Lampiris, Joseph K. Wong

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Latently infected CD4+ T cells are the main barrier to complete clearance of HIV infection, but it is unclear what mechanisms govern latent HIV infection in vivo. To address this question, we developed a new panel of reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assays specific for different HIV transcripts that define distinct blocks to transcription. We applied this panel of assays to CD4+ T cells freshly isolated from HIV-infected patients on suppressive antiretroviral therapy (ART) to quantify the degree to which different mechanisms inhibit HIV transcription. In addition, we measured the degree to which these transcriptional blocks could be reversed ex vivo by T cell activation (using anti-CD3/CD28 antibodies) or latency-reversing agents. We found that the main reversible block to HIV RNA transcription was not inhibition of transcriptional initiation but rather a series of blocks to proximal elongation, distal transcription/polyadenylation (completion), and multiple splicing. Cell dilution experiments suggested that these mechanisms operated in most of the HIV-infected CD4+ T cells examined. Latency-reversing agents exerted differential effects on the three blocks to HIV transcription, suggesting that these blocks may be governed by different mechanisms.

Original languageEnglish (US)
Article numbereaap9927
JournalScience Translational Medicine
Volume10
Issue number430
DOIs
StatePublished - Feb 28 2018
Externally publishedYes

Fingerprint

HIV
T-Lymphocytes
HIV Infections
Polyadenylation
Reverse Transcription
RNA
Polymerase Chain Reaction
Antibodies
Therapeutics

ASJC Scopus subject areas

  • Medicine(all)

Cite this

HIV latency in isolated patient CD4+ T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing. / Yukl, Steven A.; Keiser, Philip; Kim, Peggy; Telwatte, Sushama; Joshi, Sunil K.; Vu, Mai; Lampiris, Harry; Wong, Joseph K.

In: Science Translational Medicine, Vol. 10, No. 430, eaap9927, 28.02.2018.

Research output: Contribution to journalArticle

Yukl, Steven A. ; Keiser, Philip ; Kim, Peggy ; Telwatte, Sushama ; Joshi, Sunil K. ; Vu, Mai ; Lampiris, Harry ; Wong, Joseph K. / HIV latency in isolated patient CD4+ T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing. In: Science Translational Medicine. 2018 ; Vol. 10, No. 430.
@article{41026783282b4943b716a08c9cf29df1,
title = "HIV latency in isolated patient CD4+ T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing",
abstract = "Latently infected CD4+ T cells are the main barrier to complete clearance of HIV infection, but it is unclear what mechanisms govern latent HIV infection in vivo. To address this question, we developed a new panel of reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assays specific for different HIV transcripts that define distinct blocks to transcription. We applied this panel of assays to CD4+ T cells freshly isolated from HIV-infected patients on suppressive antiretroviral therapy (ART) to quantify the degree to which different mechanisms inhibit HIV transcription. In addition, we measured the degree to which these transcriptional blocks could be reversed ex vivo by T cell activation (using anti-CD3/CD28 antibodies) or latency-reversing agents. We found that the main reversible block to HIV RNA transcription was not inhibition of transcriptional initiation but rather a series of blocks to proximal elongation, distal transcription/polyadenylation (completion), and multiple splicing. Cell dilution experiments suggested that these mechanisms operated in most of the HIV-infected CD4+ T cells examined. Latency-reversing agents exerted differential effects on the three blocks to HIV transcription, suggesting that these blocks may be governed by different mechanisms.",
author = "Yukl, {Steven A.} and Philip Keiser and Peggy Kim and Sushama Telwatte and Joshi, {Sunil K.} and Mai Vu and Harry Lampiris and Wong, {Joseph K.}",
year = "2018",
month = "2",
day = "28",
doi = "10.1126/scitranslmed.aap9927",
language = "English (US)",
volume = "10",
journal = "Science Translational Medicine",
issn = "1946-6234",
publisher = "American Association for the Advancement of Science",
number = "430",

}

TY - JOUR

T1 - HIV latency in isolated patient CD4+ T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing

AU - Yukl, Steven A.

AU - Keiser, Philip

AU - Kim, Peggy

AU - Telwatte, Sushama

AU - Joshi, Sunil K.

AU - Vu, Mai

AU - Lampiris, Harry

AU - Wong, Joseph K.

PY - 2018/2/28

Y1 - 2018/2/28

N2 - Latently infected CD4+ T cells are the main barrier to complete clearance of HIV infection, but it is unclear what mechanisms govern latent HIV infection in vivo. To address this question, we developed a new panel of reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assays specific for different HIV transcripts that define distinct blocks to transcription. We applied this panel of assays to CD4+ T cells freshly isolated from HIV-infected patients on suppressive antiretroviral therapy (ART) to quantify the degree to which different mechanisms inhibit HIV transcription. In addition, we measured the degree to which these transcriptional blocks could be reversed ex vivo by T cell activation (using anti-CD3/CD28 antibodies) or latency-reversing agents. We found that the main reversible block to HIV RNA transcription was not inhibition of transcriptional initiation but rather a series of blocks to proximal elongation, distal transcription/polyadenylation (completion), and multiple splicing. Cell dilution experiments suggested that these mechanisms operated in most of the HIV-infected CD4+ T cells examined. Latency-reversing agents exerted differential effects on the three blocks to HIV transcription, suggesting that these blocks may be governed by different mechanisms.

AB - Latently infected CD4+ T cells are the main barrier to complete clearance of HIV infection, but it is unclear what mechanisms govern latent HIV infection in vivo. To address this question, we developed a new panel of reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assays specific for different HIV transcripts that define distinct blocks to transcription. We applied this panel of assays to CD4+ T cells freshly isolated from HIV-infected patients on suppressive antiretroviral therapy (ART) to quantify the degree to which different mechanisms inhibit HIV transcription. In addition, we measured the degree to which these transcriptional blocks could be reversed ex vivo by T cell activation (using anti-CD3/CD28 antibodies) or latency-reversing agents. We found that the main reversible block to HIV RNA transcription was not inhibition of transcriptional initiation but rather a series of blocks to proximal elongation, distal transcription/polyadenylation (completion), and multiple splicing. Cell dilution experiments suggested that these mechanisms operated in most of the HIV-infected CD4+ T cells examined. Latency-reversing agents exerted differential effects on the three blocks to HIV transcription, suggesting that these blocks may be governed by different mechanisms.

UR - http://www.scopus.com/inward/record.url?scp=85042848707&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042848707&partnerID=8YFLogxK

U2 - 10.1126/scitranslmed.aap9927

DO - 10.1126/scitranslmed.aap9927

M3 - Article

C2 - 29491188

AN - SCOPUS:85042848707

VL - 10

JO - Science Translational Medicine

JF - Science Translational Medicine

SN - 1946-6234

IS - 430

M1 - eaap9927

ER -