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

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 vir...

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Autores principales: Wilburn, Kaley M., Mwandumba, Henry C., Jambo, Kondwani C., Boliar, Saikat, Solouki, Sabrina, Russell, David G., Gludish, David W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Correspondence
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4982266/
https://www.ncbi.nlm.nih.gov/pubmed/27515378
http://dx.doi.org/10.1186/s12977-016-0289-2
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author Wilburn, Kaley M.
Mwandumba, Henry C.
Jambo, Kondwani C.
Boliar, Saikat
Solouki, Sabrina
Russell, David G.
Gludish, David W.
author_facet Wilburn, Kaley M.
Mwandumba, Henry C.
Jambo, Kondwani C.
Boliar, Saikat
Solouki, Sabrina
Russell, David G.
Gludish, David W.
author_sort Wilburn, Kaley M.
collection PubMed
description 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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spelling pubmed-49822662016-08-13 Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses Wilburn, Kaley M. Mwandumba, Henry C. Jambo, Kondwani C. Boliar, Saikat Solouki, Sabrina Russell, David G. Gludish, David W. Retrovirology Correspondence 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. BioMed Central 2016-08-11 /pmc/articles/PMC4982266/ /pubmed/27515378 http://dx.doi.org/10.1186/s12977-016-0289-2 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Correspondence
Wilburn, Kaley M.
Mwandumba, Henry C.
Jambo, Kondwani C.
Boliar, Saikat
Solouki, Sabrina
Russell, David G.
Gludish, David W.
Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses
title Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses
title_full Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses
title_fullStr Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses
title_full_unstemmed Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses
title_short Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses
title_sort heterogeneous loss of hiv transcription and proviral dna from 8e5/lav lymphoblastic leukemia cells revealed by rna fish:flow analyses
topic Correspondence
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4982266/
https://www.ncbi.nlm.nih.gov/pubmed/27515378
http://dx.doi.org/10.1186/s12977-016-0289-2
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