MHC–peptide tetramers for the analysis of antigen-specific T cells

Abstract

The development of the fluorescently labeled tetrameric MHC–peptide complex has enabled the direct visualization, quantification and phenotypic characterization of antigen-specific T cells using flow cytometry and has transformed our understanding of cellular immune responses. The combination of this technology with functional assays provides many new insights into these cells, allowing investigation into their lifecycle, manner of death and effector function. In this article, we hope to provide an overview of the techniques used in the construction of these tetramers, the problems and solutions associated with them, and the methods used in the study of antigen-specific T cells. Understanding how the antigen-specific cells develop and function in different circumstances and with different pathogens will be key to understanding natural host defense, as well as vaccine design and assessment.

Keywords::

• adoptive T-cell transfer
• CD4⁺ T cells
• CD8⁺ T cells
• intracellular cytokine staining
• MHC class I
• MHC class II
• phenotyping
• tetramer-guided epitope mapping
• tetramers

Financial & competing interests disclosure

This work was supported by the Wellcome Trust, the MRC, the James Martin School for the 21st Century Oxford, the NHS U19A1082830 and the NIHR Biomedical Research Centre Programme Oxford. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.