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

Abstract

Summary: 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

• Cell Biology
• Cell culture
• Cell isolation
• Flow Cytometry/Mass Cytometry
• Cell-based Assays
• Immunology