Characterization of Mouse γδ T Cell Subsets in the Setting of Type-2 Immunity

Methods Mol Biol. 2018:1799:135-151. doi: 10.1007/978-1-4939-7896-0_12.

Abstract

Accumulating evidence indicates that γδ T cells are a critical component of type-2 immunity. However, the role of these cells in type-2 immune responses seems to be divergent. γδ T cells are heterogeneous lymphocytes that can be further divided into TCR-Vγ/δ definable subsets. Different subsets have distinct and sometimes opposite function during immune responses. In this chapter, we describe the detailed protocol for characterization of γδ T cell subsets in a mouse model of ovalbumin (OVA)/alum-induced type-2 immunity. Our protocol includes identifying γδ T cell subsets by flow cytometry, functionally inactivating individual subsets in vivo, purifying γδ T cell subsets, and using adoptive cell transfer to explore the role of individual subsets in OVA/alum-induced IgE responses.

Keywords: Adoptive cell transfer; Flow cytometry; IgE; OVA/alum; Subsets; TCR-Vγ/δ; Type-2 immunity; γδ T cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer
  • Alum Compounds
  • Animals
  • Antibodies, Monoclonal / immunology
  • Biomarkers
  • Cell Separation / methods
  • Flow Cytometry
  • Immune Tolerance
  • Immunity, Cellular*
  • Immunoglobulin E / immunology
  • Immunophenotyping
  • Lung / immunology
  • Lung / metabolism
  • Mice
  • Mice, Knockout
  • Mice, Transgenic
  • Ovalbumin / immunology
  • Receptors, Antigen, T-Cell, gamma-delta / genetics
  • Receptors, Antigen, T-Cell, gamma-delta / metabolism*
  • Spleen / immunology
  • Spleen / metabolism
  • T-Lymphocyte Subsets / cytology
  • T-Lymphocyte Subsets / immunology*
  • T-Lymphocyte Subsets / metabolism*

Substances

  • Alum Compounds
  • Antibodies, Monoclonal
  • Biomarkers
  • Receptors, Antigen, T-Cell, gamma-delta
  • ovalbumin-alum
  • Immunoglobulin E
  • Ovalbumin