In Situ Peptide-MHC-II Tetramer Staining of Antigen-Specific CD4+ T Cells in Tissues |
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| Authors: | Thamotharampillai Dileepan Hyeon O Kim P Patrick Cleary Pamela J Skinner |
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| Institution: | 1. Department of Microbiology, University of Minnesota, Minneapolis, MN, United States of America.; 2. Department of Veterinary and Biomedical Sciences, University of Minnesota, Saint Paul, MN, United States of America.; Johns Hopkins University, UNITED STATES, |
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| Abstract: | The invention of peptide-MHC-tetramer technology to label antigen-specific T cells has led to an enhanced understanding of T lymphocyte biology. Here we describe the development of an in situ pMHC-II tetramer staining method to visualize antigen-specific CD4+ T cells in tissues. This method complements other methods developed that similarly use MHC class II reagents to stain antigen-specific CD4+ T cells in situ. In this study, we used group A streptococcus (GAS) expressing a surrogate peptide (2W) to inoculate C57BL/6 mice, and used fresh nasal-associated lymphoid tissues (NALT) in optimizing the in situ staining of 2W:I-Ab specific CD4+ T cells. The results showed 2W:I-Ab tetramer-binding CD4+ T cells in GAS-2W but not GAS infected mice. This method holds promise to be broadly applicable to study the localization, abundance, and phenotype of antigen-specific CD4+ T cells in undisrupted tissues. |
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