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Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining
MHC tetramers are an essential tool for characterizing antigen-specific CD4+ T cells. However, their ex vivo analysis is limited by the large sample requirements. Here we demonstrate a combinatorial staining approach that allows simultaneous characterization of multiple specificities to address this...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5013714/ https://www.ncbi.nlm.nih.gov/pubmed/27571776 http://dx.doi.org/10.1038/ncomms12614 |
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author | Uchtenhagen, Hannes Rims, Cliff Blahnik, Gabriele Chow, I-Ting Kwok, William W. Buckner, Jane H. James, Eddie A. |
author_facet | Uchtenhagen, Hannes Rims, Cliff Blahnik, Gabriele Chow, I-Ting Kwok, William W. Buckner, Jane H. James, Eddie A. |
author_sort | Uchtenhagen, Hannes |
collection | PubMed |
description | MHC tetramers are an essential tool for characterizing antigen-specific CD4+ T cells. However, their ex vivo analysis is limited by the large sample requirements. Here we demonstrate a combinatorial staining approach that allows simultaneous characterization of multiple specificities to address this challenge. As proof of principle, we analyse CD4+ T-cell responses to the seasonal influenza vaccine, establishing a frequency hierarchy and examining differences in memory and activation status, lineage commitment and cytokine expression. We also observe cross-reactivity between an established epitope and recent variant and provide a means for probing T-cell receptor cross-reactivity. Using cord blood samples, we correlate the adult frequency hierarchy with the naive precursor frequencies. Last, we use our combinatorial staining approach to demonstrate that rheumatoid arthritis patients on therapy can mount effective responses to influenza vaccination. Together, these results demonstrate the utility of combinatorial tetramer staining and suggest that this approach may have broad applicability in human health and disease. |
format | Online Article Text |
id | pubmed-5013714 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-50137142016-09-20 Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining Uchtenhagen, Hannes Rims, Cliff Blahnik, Gabriele Chow, I-Ting Kwok, William W. Buckner, Jane H. James, Eddie A. Nat Commun Article MHC tetramers are an essential tool for characterizing antigen-specific CD4+ T cells. However, their ex vivo analysis is limited by the large sample requirements. Here we demonstrate a combinatorial staining approach that allows simultaneous characterization of multiple specificities to address this challenge. As proof of principle, we analyse CD4+ T-cell responses to the seasonal influenza vaccine, establishing a frequency hierarchy and examining differences in memory and activation status, lineage commitment and cytokine expression. We also observe cross-reactivity between an established epitope and recent variant and provide a means for probing T-cell receptor cross-reactivity. Using cord blood samples, we correlate the adult frequency hierarchy with the naive precursor frequencies. Last, we use our combinatorial staining approach to demonstrate that rheumatoid arthritis patients on therapy can mount effective responses to influenza vaccination. Together, these results demonstrate the utility of combinatorial tetramer staining and suggest that this approach may have broad applicability in human health and disease. Nature Publishing Group 2016-08-30 /pmc/articles/PMC5013714/ /pubmed/27571776 http://dx.doi.org/10.1038/ncomms12614 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Uchtenhagen, Hannes Rims, Cliff Blahnik, Gabriele Chow, I-Ting Kwok, William W. Buckner, Jane H. James, Eddie A. Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining |
title | Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining |
title_full | Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining |
title_fullStr | Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining |
title_full_unstemmed | Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining |
title_short | Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining |
title_sort | efficient ex vivo analysis of cd4+ t-cell responses using combinatorial hla class ii tetramer staining |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5013714/ https://www.ncbi.nlm.nih.gov/pubmed/27571776 http://dx.doi.org/10.1038/ncomms12614 |
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