Cargando…

Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status

RNA flow cytometry (FISH-Flow) achieves high-throughput measurement of single-cell gene expression by combining in-situ nucleic acid hybridization with flow cytometry. We tested whether antigen-specific T-cell responses detected by FISH-Flow correlated with latent tuberculosis infection (LTBI), a co...

Descripción completa

Detalles Bibliográficos
Autores principales: Vir, Pooja, Arrigucci, Riccardo, Lakehal, Karim, Davidow, Amy L., Pine, Richard, Tyagi, Sanjay, Bushkin, Yuri, Lardizabal, Alfred, Gennaro, Maria Laura
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4681842/
https://www.ncbi.nlm.nih.gov/pubmed/26658491
http://dx.doi.org/10.1371/journal.pone.0144904
_version_ 1782405782957981696
author Vir, Pooja
Arrigucci, Riccardo
Lakehal, Karim
Davidow, Amy L.
Pine, Richard
Tyagi, Sanjay
Bushkin, Yuri
Lardizabal, Alfred
Gennaro, Maria Laura
author_facet Vir, Pooja
Arrigucci, Riccardo
Lakehal, Karim
Davidow, Amy L.
Pine, Richard
Tyagi, Sanjay
Bushkin, Yuri
Lardizabal, Alfred
Gennaro, Maria Laura
author_sort Vir, Pooja
collection PubMed
description RNA flow cytometry (FISH-Flow) achieves high-throughput measurement of single-cell gene expression by combining in-situ nucleic acid hybridization with flow cytometry. We tested whether antigen-specific T-cell responses detected by FISH-Flow correlated with latent tuberculosis infection (LTBI), a condition affecting one-third of the world population. Peripheral-blood mononuclear cells from donors, identified as positive or negative for LTBI by current medical practice, were stimulated ex vivo with mycobacterial antigen. IFNG and IL2 mRNA production was assayed by FISH-Flow. Concurrently, immunophenotypes of the cytokine mRNA-positive cells were characterized by conventional, antibody-based staining of cell-surface markers. An association was found between donor LTBI status and antigen-specific induction of IFNG and IL2 transcripts. Induction of these cytokine genes, which was detected by FISH-Flow in a quarter the time required to see release of the corresponding proteins by ELISA, occurred primarily in activated CD4+ T cells via T-cell receptor engagement. Moreover, NK cells contributed to IFNG gene induction. These results show that antigen-driven induction of T-cell cytokine mRNA is a measurable single-cell parameter of the host responses associated with latent tuberculosis. FISH-Flow read-outs contribute a multi-scale dimension to the immunophenotyping afforded by antibody-based flow cytometry. Multi-scale, single-cell analyses may satisfy the need to determine disease stage and therapy response for tuberculosis and other infectious pathologies.
format Online
Article
Text
id pubmed-4681842
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-46818422015-12-31 Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status Vir, Pooja Arrigucci, Riccardo Lakehal, Karim Davidow, Amy L. Pine, Richard Tyagi, Sanjay Bushkin, Yuri Lardizabal, Alfred Gennaro, Maria Laura PLoS One Research Article RNA flow cytometry (FISH-Flow) achieves high-throughput measurement of single-cell gene expression by combining in-situ nucleic acid hybridization with flow cytometry. We tested whether antigen-specific T-cell responses detected by FISH-Flow correlated with latent tuberculosis infection (LTBI), a condition affecting one-third of the world population. Peripheral-blood mononuclear cells from donors, identified as positive or negative for LTBI by current medical practice, were stimulated ex vivo with mycobacterial antigen. IFNG and IL2 mRNA production was assayed by FISH-Flow. Concurrently, immunophenotypes of the cytokine mRNA-positive cells were characterized by conventional, antibody-based staining of cell-surface markers. An association was found between donor LTBI status and antigen-specific induction of IFNG and IL2 transcripts. Induction of these cytokine genes, which was detected by FISH-Flow in a quarter the time required to see release of the corresponding proteins by ELISA, occurred primarily in activated CD4+ T cells via T-cell receptor engagement. Moreover, NK cells contributed to IFNG gene induction. These results show that antigen-driven induction of T-cell cytokine mRNA is a measurable single-cell parameter of the host responses associated with latent tuberculosis. FISH-Flow read-outs contribute a multi-scale dimension to the immunophenotyping afforded by antibody-based flow cytometry. Multi-scale, single-cell analyses may satisfy the need to determine disease stage and therapy response for tuberculosis and other infectious pathologies. Public Library of Science 2015-12-14 /pmc/articles/PMC4681842/ /pubmed/26658491 http://dx.doi.org/10.1371/journal.pone.0144904 Text en © 2015 Vir et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Vir, Pooja
Arrigucci, Riccardo
Lakehal, Karim
Davidow, Amy L.
Pine, Richard
Tyagi, Sanjay
Bushkin, Yuri
Lardizabal, Alfred
Gennaro, Maria Laura
Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status
title Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status
title_full Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status
title_fullStr Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status
title_full_unstemmed Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status
title_short Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status
title_sort single-cell cytokine gene expression in peripheral blood cells correlates with latent tuberculosis status
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4681842/
https://www.ncbi.nlm.nih.gov/pubmed/26658491
http://dx.doi.org/10.1371/journal.pone.0144904
work_keys_str_mv AT virpooja singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT arrigucciriccardo singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT lakehalkarim singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT davidowamyl singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT pinerichard singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT tyagisanjay singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT bushkinyuri singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT lardizabalalfred singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus
AT gennaromarialaura singlecellcytokinegeneexpressioninperipheralbloodcellscorrelateswithlatenttuberculosisstatus