Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses

Kaley M. Wilburn; Henry Mwandumba; Kondwani Jambo; Saikat Boliar; Sabrina Solouki; David G. Russell; David W. Gludish

doi:10.1186/s12977-016-0289-2

Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses

Kaley M. Wilburn, Henry Mwandumba, Kondwani Jambo, Saikat Boliar, Sabrina Solouki, David G. Russell, David W. Gludish

Clinical Sciences

Research output: Contribution to journal › Article › peer-review

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Abstract

8E5/LAV cells harbor a single HIV provirus, and are used frequently to generate standards for HIV genome quantification. Using flow cytometry-based in situ mRNA hybridization validated by qPCR, we find that different batches of 8E5 cells contain varying numbers of cells lacking viral mRNA and/or viral genomes. These findings raise concerns for studies employing 8E5 cells for quantitation, and highlight the value of mRNA FISH and flow cytometry in the detection and enumeration of HIV-positive cells.

Original language	English
Article number	55
Journal	Retrovirology
Volume	13
Issue number	1
DOIs	https://doi.org/10.1186/s12977-016-0289-2
Publication status	Published - 11 Aug 2016

Keywords

8E5
FISH:FACS
FISH:FLOW
HIV cell-associated RNA
HIV flow cytometry assay
HIV proviral loss
HIV proviral silencing
HIV RNA in situ hybridization
Reservoir quantitation

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Retrovirology 13 55 Heterogenous loss of HIV transcriptionFinal published version, 1.47 MBLicence: CC BY

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title = "Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses",

abstract = "8E5/LAV cells harbor a single HIV provirus, and are used frequently to generate standards for HIV genome quantification. Using flow cytometry-based in situ mRNA hybridization validated by qPCR, we find that different batches of 8E5 cells contain varying numbers of cells lacking viral mRNA and/or viral genomes. These findings raise concerns for studies employing 8E5 cells for quantitation, and highlight the value of mRNA FISH and flow cytometry in the detection and enumeration of HIV-positive cells.",

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day = "11",

doi = "10.1186/s12977-016-0289-2",

language = "English",

volume = "13",

journal = "Retrovirology",

issn = "1742-4690",

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T1 - Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses

AU - Wilburn, Kaley M.

AU - Mwandumba, Henry

AU - Jambo, Kondwani

AU - Boliar, Saikat

AU - Solouki, Sabrina

AU - Russell, David G.

AU - Gludish, David W.

PY - 2016/8/11

Y1 - 2016/8/11

N2 - 8E5/LAV cells harbor a single HIV provirus, and are used frequently to generate standards for HIV genome quantification. Using flow cytometry-based in situ mRNA hybridization validated by qPCR, we find that different batches of 8E5 cells contain varying numbers of cells lacking viral mRNA and/or viral genomes. These findings raise concerns for studies employing 8E5 cells for quantitation, and highlight the value of mRNA FISH and flow cytometry in the detection and enumeration of HIV-positive cells.

AB - 8E5/LAV cells harbor a single HIV provirus, and are used frequently to generate standards for HIV genome quantification. Using flow cytometry-based in situ mRNA hybridization validated by qPCR, we find that different batches of 8E5 cells contain varying numbers of cells lacking viral mRNA and/or viral genomes. These findings raise concerns for studies employing 8E5 cells for quantitation, and highlight the value of mRNA FISH and flow cytometry in the detection and enumeration of HIV-positive cells.

KW - 8E5

KW - FISH:FACS

KW - FISH:FLOW

KW - HIV cell-associated RNA

KW - HIV flow cytometry assay

KW - HIV proviral loss

KW - HIV proviral silencing

KW - HIV RNA in situ hybridization

KW - Reservoir quantitation

U2 - 10.1186/s12977-016-0289-2

DO - 10.1186/s12977-016-0289-2

M3 - Article

SN - 1742-4690

VL - 13

JO - Retrovirology

JF - Retrovirology

IS - 1

M1 - 55

ER -

Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses

Abstract

Keywords

UN SDGs

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