In situ MHC-tetramer staining and quantitative analysis to determine the location, abundance, and phenotype of antigen-specific CD8 T cells in tissues

Shengbin Li, Gwantwa Mwakalundwa, Pamela J Skinner

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

T cells are critical to many immunological processes, including detecting and eliminating virus-infected cells, preventing autoimmunity, assisting in B-cell and plasma-cell production of antibodies, and detecting and eliminating cancer cells. The development of MHC-tetramer staining of antigen-specific T cells analyzed by flow cytometry has revolutionized our ability to study and understand the immunobiology of T cells. While extremely useful for determining the quantity and phenotype of antigen-specific T cells, flow cytometry cannot determine the spatial localization of antigen-specific T cells to other cells and structures in tissues, and current disaggregation techniques to extract the T cells needed for flow cytometry have limited effectiveness in non-lymphoid tissues. In situ MHC-tetramer staining (IST) is a technique to visualize T cells that are specific for antigens of interest in tissues. In combination with immunohistochemistry (IHC), IST can determine the abundance, location, and phenotype of antigen-specific CD8 and CD4 T cells in tissues. Here, we describe a protocol to stain and enumerate antigen-specific CD8 T cells, with specific phenotypes located within specific tissue compartments. These procedures are the same that we used in our recent publication by Li et al., entitled "Simian Immunodeficiency Virus-Producing Cells in Follicles Are Partially Suppressed by CD8+ Cells In Vivo." The methods described are broadly applicable because they can be used to localize, phenotype, and quantify essentially any antigen-specific CD8 T cell for which MHC tetramers are available, in any tissue.

Original languageEnglish (US)
Article numbere56130
JournalJournal of Visualized Experiments
Volume2017
Issue number127
DOIs
StatePublished - Sep 22 2017

Bibliographical note

Funding Information:
This work was supported by Public Health Service grants from the National Institutes of Health (T32 DA007097, R01AI096966, andUM1AI26617).

Keywords

  • Confocal microscopy
  • Immunohistochemistry
  • Immunology
  • In situ tetramer staining (IST)
  • Issue 127
  • Localization
  • Phenotype
  • Quantitative image analysis
  • Virus-specific CD8+ T cells

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