Efficient ex vivo analysis of CD4+ T-cell responses using combinatorial HLA class II tetramer staining

Hannes Uchtenhagen, Cliff Rims, Gabriele Blahnik, I-Ting Chow, William W. Kwok, Jane H. Buckner and Eddie A. James ()
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Hannes Uchtenhagen: Benaroya Research Institute at Virginia Mason, Translational Research Program
Cliff Rims: Benaroya Research Institute at Virginia Mason, Translational Research Program
Gabriele Blahnik: Benaroya Research Institute at Virginia Mason, Diabetes Program
I-Ting Chow: Benaroya Research Institute at Virginia Mason, Diabetes Program
William W. Kwok: Benaroya Research Institute at Virginia Mason, Diabetes Program
Jane H. Buckner: Benaroya Research Institute at Virginia Mason, Translational Research Program
Eddie A. James: Benaroya Research Institute at Virginia Mason, Diabetes Program and Tetramer Core Laboratory

Nature Communications, 2016, vol. 7, issue 1, 1-12

Abstract: Abstract 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.

Date: 2016
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DOI: 10.1038/ncomms12614

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