title: In Situ Peptide-MHC-II Tetramer Staining of Antigen-Specific CD4+ T Cells in Tissues.
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.
authors.affiliation: Department of Microbiology, University of Minnesota, Minneapolis, MN, United States of America.
authors.forename: Thamotharampillai
authors.initials: T
authors.lastname: Dileepan
authors.affiliation: Department of Veterinary and Biomedical Sciences, University of Minnesota, Saint Paul, MN, United States of America.
authors.forename: Hyeon O
authors.initials: HO
authors.lastname: Kim
authors.affiliation: Department of Microbiology, University of Minnesota, Minneapolis, MN, United States of America.
authors.forename: P Patrick
authors.initials: PP
authors.lastname: Cleary
authors.affiliation: Department of Veterinary and Biomedical Sciences, University of Minnesota, Saint Paul, MN, United States of America.
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Skinner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D013290:Streptococcal Infections / Q000382:microbiology / Q000473:pathology / Q000662:veterinary; D013297:Streptococcus pyogenes / Q000378:metabolism / Q000502:physiology
chemical_list: D018984:Epitopes, T-Lymphocyte; D010455:Peptides; C011080:polyglycine