Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays

STAR Protoc. 2021 Nov 16;2(4):100961. doi: 10.1016/j.xpro.2021.100961. eCollection 2021 Dec 17.

Abstract

Specific antigen recognition by T cell receptor (TCR) activates TCR signaling pathway, leading to T cell proliferation and differentiation into effector and memory cells. Herein, we describe protocols for TCR stimulation assays, including procedures for the isolation and enrichment of mouse splenic T cells for ex vivo TCR stimulation with anti-CD3/CD28 antibodies, and the use of ovalbumin-OT-II mouse model for in vivo TCR stimulation. We applied this protocol to show that MYC protein is essential for T cell proliferation and differentiation. For complete details on the use and execution of this protocol, please refer to Nozais et al. (2021).

Keywords: Antibody; Cell Biology; Cell culture; Cell isolation; Cell-based Assays; Flow Cytometry/Mass Cytometry; Immunology; Model Organisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Separation / methods*
  • Female
  • Immunologic Tests
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Receptors, Antigen, T-Cell* / immunology
  • Receptors, Antigen, T-Cell* / metabolism
  • Spleen / cytology*
  • T-Lymphocytes* / cytology
  • T-Lymphocytes* / immunology
  • T-Lymphocytes* / metabolism

Substances

  • Receptors, Antigen, T-Cell