Generation of MR1-Restricted T Cell Clones by Limiting Dilution Cloning of MR1 Tetramer+ Cells

Methods Mol Biol. 2020:2098:219-235. doi: 10.1007/978-1-0716-0207-2_15.

Abstract

Tetramers are a powerful tool for identification of T cell subsets that are restricted by specific antigen presenting molecules and their cognate antigens. The generation of T cell clones from specific T cell subsets allows for further investigation of the phenotype and function of these cells. Here, we describe a method for sorting and cloning of MR1-restricted T cells using the MR1/5-OP-RU tetramer. This protocol can be easily modified to enrich for expansion of specific or unique subsets of MR1-restricted T cell clones from any tissue to further characterize the phenotype and function of those cells.

Keywords: MAIT cells; MR1-restricted T cells; T cell cloning.

MeSH terms

  • Biomarkers
  • Cell Culture Techniques
  • Cells, Cultured
  • Clone Cells*
  • Cytokines / metabolism
  • Enzyme-Linked Immunospot Assay
  • Flow Cytometry
  • Histocompatibility Antigens Class I / chemistry
  • Histocompatibility Antigens Class I / metabolism*
  • Humans
  • Immunophenotyping
  • Minor Histocompatibility Antigens / chemistry
  • Minor Histocompatibility Antigens / metabolism*
  • Phenotype
  • Protein Multimerization*
  • T-Lymphocyte Subsets / immunology*
  • T-Lymphocyte Subsets / metabolism*

Substances

  • Biomarkers
  • Cytokines
  • Histocompatibility Antigens Class I
  • MR1 protein, human
  • Minor Histocompatibility Antigens