Immune Monitoring of Human Mucosal-Associated Invariant T Cells by Quantitative Proteomics

Methods Mol Biol. 2020:2098:209-218. doi: 10.1007/978-1-0716-0207-2_14.

Abstract

Molecular phenotypes of mucosal-associated invariant T (MAIT) cells are correlating with individual susceptibilities and outcomes in human diseases. Quantitative proteome strategies can examine such variations in the functional and druggable inventory of MAIT cells comprehensively, but protocols for the support of translational and clinical studies are still rare. Here, we describe a protocol in which MR1-restricted MAIT cells were isolated from blood donations by FACS and are then characterized by quantitative proteomics (iTRAQ-LC-MS/MS) to complement information about their unique effector phenotype and to investigate donor-/patient- or disease-specific variations in protein networks with high precision.

Keywords: 8-plex iTRAQ™; Clinical proteomics; MR1 tetramer; Mucosal-associated invariant T cells.

MeSH terms

  • Biomarkers
  • Cell Separation / methods
  • Chromatography, Liquid
  • Histocompatibility Antigens Class I / metabolism
  • Humans
  • Mass Spectrometry
  • Minor Histocompatibility Antigens / metabolism
  • Monitoring, Immunologic* / methods
  • Mucosal-Associated Invariant T Cells / immunology*
  • Mucosal-Associated Invariant T Cells / metabolism*
  • Proteome*
  • Proteomics* / methods
  • Staining and Labeling
  • Tandem Mass Spectrometry

Substances

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