Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs

Major histocompatibility complex (MHC) tetramers can work as diagnostic tools to identify antigen-specific T cells in immunological research and monitoring. Here, we provide a general protocol for the production of MHC tetramer. We obtain highly pure N-terminal His-tagged HLA-A2 α chain and β2-micro...

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Autores principales: Ye, Jiezhou, Lei, Wen, Xiao, ChanChan, Su, Xiangmeng, Gao, Jie, Li, Xinran, Li, Wenxin, Wang, Pengcheng, Jiang, Zhenyou, Chen, Guobing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914994/
https://www.ncbi.nlm.nih.gov/pubmed/35284834
http://dx.doi.org/10.1016/j.xpro.2022.101206
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author Ye, Jiezhou
Lei, Wen
Xiao, ChanChan
Su, Xiangmeng
Gao, Jie
Li, Xinran
Li, Wenxin
Wang, Pengcheng
Jiang, Zhenyou
Chen, Guobing
author_facet Ye, Jiezhou
Lei, Wen
Xiao, ChanChan
Su, Xiangmeng
Gao, Jie
Li, Xinran
Li, Wenxin
Wang, Pengcheng
Jiang, Zhenyou
Chen, Guobing
author_sort Ye, Jiezhou
collection PubMed
description Major histocompatibility complex (MHC) tetramers can work as diagnostic tools to identify antigen-specific T cells in immunological research and monitoring. Here, we provide a general protocol for the production of MHC tetramer. We obtain highly pure N-terminal His-tagged HLA-A2 α chain and β2-microglobulin (β(2)m) to fold a monomer with a photocleavable peptide, which can exchange with an HLA-A2 presented peptide derived from influenza A virus. Further those monomers compose tetramer to stain antigen-specific CD8(+) T cells. For complete details on the use and execution of this protocol, please refer to Xiao C.C. et al. (2021).
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spelling pubmed-89149942022-03-12 Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs Ye, Jiezhou Lei, Wen Xiao, ChanChan Su, Xiangmeng Gao, Jie Li, Xinran Li, Wenxin Wang, Pengcheng Jiang, Zhenyou Chen, Guobing STAR Protoc Protocol Major histocompatibility complex (MHC) tetramers can work as diagnostic tools to identify antigen-specific T cells in immunological research and monitoring. Here, we provide a general protocol for the production of MHC tetramer. We obtain highly pure N-terminal His-tagged HLA-A2 α chain and β2-microglobulin (β(2)m) to fold a monomer with a photocleavable peptide, which can exchange with an HLA-A2 presented peptide derived from influenza A virus. Further those monomers compose tetramer to stain antigen-specific CD8(+) T cells. For complete details on the use and execution of this protocol, please refer to Xiao C.C. et al. (2021). Elsevier 2022-03-09 /pmc/articles/PMC8914994/ /pubmed/35284834 http://dx.doi.org/10.1016/j.xpro.2022.101206 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ye, Jiezhou
Lei, Wen
Xiao, ChanChan
Su, Xiangmeng
Gao, Jie
Li, Xinran
Li, Wenxin
Wang, Pengcheng
Jiang, Zhenyou
Chen, Guobing
Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs
title Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs
title_full Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs
title_fullStr Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs
title_full_unstemmed Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs
title_short Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs
title_sort production and purification of infectious diseases-associated tetramer for staining of cd8(+) t cells from pbmcs
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914994/
https://www.ncbi.nlm.nih.gov/pubmed/35284834
http://dx.doi.org/10.1016/j.xpro.2022.101206
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