Gene Expression-Based Identification of Antigen-Responsive CD8+ T Cells on a Single-Cell Level

Front Immunol. 2019 Nov 6:10:2568. doi: 10.3389/fimmu.2019.02568. eCollection 2019.

Abstract

CD8+ T cells are important effectors of adaptive immunity against pathogens, tumors, and self antigens. Here, we asked how human cognate antigen-responsive CD8+ T cells and their receptors could be identified in unselected single-cell gene expression data. Single-cell RNA sequencing and qPCR of dye-labeled antigen-specific cells identified large gene sets that were congruently up- or downregulated in virus-responsive CD8+ T cells under different antigen presentation conditions. Combined expression of TNFRSF9, XCL1, XCL2, and CRTAM was the most distinct marker of virus-responsive cells on a single-cell level. Using transcriptomic data, we developed a machine learning-based classifier that provides sensitive and specific detection of virus-responsive CD8+ T cells from unselected populations. Gene response profiles of CD8+ T cells specific for the autoantigen islet-specific glucose-6-phosphatase catalytic subunit-related protein differed markedly from virus-specific cells. These findings provide single-cell gene expression parameters for comprehensive identification of rare antigen-responsive cells and T cell receptors.

Keywords: CD8+ T cells; CMV pp65; CTL (cytotoxic T lymphocyte); T cell receptor (TCR); antigen-responsive; gene-expression analysis; influenza matrix protein; single-cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen Presentation / immunology
  • Antigen-Presenting Cells / immunology
  • Antigen-Presenting Cells / metabolism
  • Antigens / immunology*
  • Autoantigens / immunology
  • CD8-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / metabolism*
  • Gene Expression Profiling* / methods
  • Histocompatibility Antigens Class I / chemistry
  • Histocompatibility Antigens Class I / immunology
  • Histocompatibility Antigens Class I / metabolism
  • Humans
  • Peptides / immunology
  • Single-Cell Analysis* / methods
  • Viral Matrix Proteins / chemistry
  • Viral Matrix Proteins / immunology

Substances

  • Antigens
  • Autoantigens
  • Histocompatibility Antigens Class I
  • M1 protein, Influenza A virus
  • Peptides
  • Viral Matrix Proteins
  • cytomegalovirus matrix protein 65kDa