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Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes

The cytotoxicity of epitope-specific CD8(+) T cells is usually measured indirectly through IFNγ production. Existing assays that directly measure this activity are limited mainly to measurements of up to two specificities in a single reaction. Here, we develop a multiplex cytotoxicity assay that all...

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Autores principales: Poh, Chek Meng, Zheng, Jian, Channappanavar, Rudragouda, Chang, Zi Wei, Nguyen, Thi H. O., Rénia, Laurent, Kedzierska, Katherine, Perlman, Stanley, Poon, Leo L. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7078160/
https://www.ncbi.nlm.nih.gov/pubmed/32218786
http://dx.doi.org/10.3389/fimmu.2020.00400
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author Poh, Chek Meng
Zheng, Jian
Channappanavar, Rudragouda
Chang, Zi Wei
Nguyen, Thi H. O.
Rénia, Laurent
Kedzierska, Katherine
Perlman, Stanley
Poon, Leo L. M.
author_facet Poh, Chek Meng
Zheng, Jian
Channappanavar, Rudragouda
Chang, Zi Wei
Nguyen, Thi H. O.
Rénia, Laurent
Kedzierska, Katherine
Perlman, Stanley
Poon, Leo L. M.
author_sort Poh, Chek Meng
collection PubMed
description The cytotoxicity of epitope-specific CD8(+) T cells is usually measured indirectly through IFNγ production. Existing assays that directly measure this activity are limited mainly to measurements of up to two specificities in a single reaction. Here, we develop a multiplex cytotoxicity assay that allows direct, simultaneous measurement of up to 23 different specificities of CD8(+) T cells in a single reaction. This can greatly reduce the amount of starting clinical materials for a systematic screening of CD8(+) T cell epitopes. In addition, this greatly enhanced capacity enables the incorporation of irrelevant epitopes for determining the non-specific killing activity of CD8(+) T cells, thereby allowing to measure the actual epitope-specific cytotoxicity activities. This technique is shown to be useful to study both human and mouse CD8(+) T cells. Besides, our results from human PBMCs and three independent infectious animal models (MERS, influenza and malaria) further reveal that IFNγ expression by epitope-specific CD8(+) T cells does not always correlate with their cell-killing potential, highlighting the need for using cytotoxicity assays in specific contexts (e.g., evaluating vaccine candidates). Overall, our approach opens up new possibilities for comprehensive analyses of CD8(+) T cell cytotoxicity in a practical manner.
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spelling pubmed-70781602020-03-26 Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes Poh, Chek Meng Zheng, Jian Channappanavar, Rudragouda Chang, Zi Wei Nguyen, Thi H. O. Rénia, Laurent Kedzierska, Katherine Perlman, Stanley Poon, Leo L. M. Front Immunol Immunology The cytotoxicity of epitope-specific CD8(+) T cells is usually measured indirectly through IFNγ production. Existing assays that directly measure this activity are limited mainly to measurements of up to two specificities in a single reaction. Here, we develop a multiplex cytotoxicity assay that allows direct, simultaneous measurement of up to 23 different specificities of CD8(+) T cells in a single reaction. This can greatly reduce the amount of starting clinical materials for a systematic screening of CD8(+) T cell epitopes. In addition, this greatly enhanced capacity enables the incorporation of irrelevant epitopes for determining the non-specific killing activity of CD8(+) T cells, thereby allowing to measure the actual epitope-specific cytotoxicity activities. This technique is shown to be useful to study both human and mouse CD8(+) T cells. Besides, our results from human PBMCs and three independent infectious animal models (MERS, influenza and malaria) further reveal that IFNγ expression by epitope-specific CD8(+) T cells does not always correlate with their cell-killing potential, highlighting the need for using cytotoxicity assays in specific contexts (e.g., evaluating vaccine candidates). Overall, our approach opens up new possibilities for comprehensive analyses of CD8(+) T cell cytotoxicity in a practical manner. Frontiers Media S.A. 2020-03-11 /pmc/articles/PMC7078160/ /pubmed/32218786 http://dx.doi.org/10.3389/fimmu.2020.00400 Text en Copyright © 2020 Poh, Zheng, Channappanavar, Chang, Nguyen, Rénia, Kedzierska, Perlman and Poon. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Poh, Chek Meng
Zheng, Jian
Channappanavar, Rudragouda
Chang, Zi Wei
Nguyen, Thi H. O.
Rénia, Laurent
Kedzierska, Katherine
Perlman, Stanley
Poon, Leo L. M.
Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes
title Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes
title_full Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes
title_fullStr Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes
title_full_unstemmed Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes
title_short Multiplex Screening Assay for Identifying Cytotoxic CD8(+) T Cell Epitopes
title_sort multiplex screening assay for identifying cytotoxic cd8(+) t cell epitopes
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7078160/
https://www.ncbi.nlm.nih.gov/pubmed/32218786
http://dx.doi.org/10.3389/fimmu.2020.00400
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