Journal article
Multiplex Screening Assay for Identifying Cytotoxic CD8 + T Cell Epitopes
Frontiers in immunology, Vol.11, pp.400-400
2020
DOI: 10.3389/fimmu.2020.00400
PMCID: PMC7078160
PMID: 32218786
Abstract
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.
Details
- Title: Subtitle
- Multiplex Screening Assay for Identifying Cytotoxic CD8 + T Cell Epitopes
- Creators
- Chek Meng Poh - School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, ChinaJian Zheng - Department of Microbiology and Immunology, Carver College of Medicine, University of Iowa, Iowa City, IA, United StatesRudragouda Channappanavar - Department of Microbiology and Immunology, Carver College of Medicine, University of Iowa, Iowa City, IA, United StatesZi Wei Chang - Singapore Immunology Network, Agency of Science, Technology and Research, Singapore, SingaporeThi H O Nguyen - Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, AustraliaLaurent Rénia - Singapore Immunology Network, Agency of Science, Technology and Research, Singapore, SingaporeKatherine Kedzierska - Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, AustraliaStanley Perlman - Department of Microbiology and Immunology, Carver College of Medicine, University of Iowa, Iowa City, IA, United StatesLeo L M Poon - School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
- Resource Type
- Journal article
- Publication Details
- Frontiers in immunology, Vol.11, pp.400-400
- DOI
- 10.3389/fimmu.2020.00400
- PMID
- 32218786
- PMCID
- PMC7078160
- NLM abbreviation
- Front Immunol
- ISSN
- 1664-3224
- eISSN
- 1664-3224
- Publisher
- Switzerland
- Grant note
- P01 AI060699 / NIAID NIH HHS R01 AI129269 / NIAID NIH HHS HHSN272201400006C / NIAID NIH HHS
- Language
- English
- Date published
- 2020
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Infectious Disease (Pediatrics)
- Record Identifier
- 9984070614702771
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