Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Uchtenhagen, Hannes, Rims, Cliff, Blahnik, Gabriele, Chow, I-Ting, Kwok, William W., Buckner, Jane H., James, Eddie A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
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
_version_ 1782452219627438080
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
work_keys_str_mv AT uchtenhagenhannes efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining
AT rimscliff efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining
AT blahnikgabriele efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining
AT chowiting efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining
AT kwokwilliamw efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining
AT bucknerjaneh efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining
AT jameseddiea efficientexvivoanalysisofcd4tcellresponsesusingcombinatorialhlaclassiitetramerstaining