Evaluation of in vivo target cell elimination by antigen-specific cytotoxic T lymphocytes

Methods Cell Biol. 2025:197:141-155. doi: 10.1016/bs.mcb.2025.03.025. Epub 2025 May 3.

Abstract

The pivotal role of cytotoxic T lymphocyte (CTL) killing of target cells in vivo continues to be underscored by emerging research. CTLs are antigen-specific effector CD8 + T lymphocytes that serve as adaptive defenders against a myriad of threats, including viral infections, cancerous cells, and other pathogenic invaders. In vivo CTL killing assays contemplate the interaction of effector and target cells in the context of a proper microenvironment, making the analysis biologically more relevant than in vitro assays. They play a crucial role in advancing our understanding of immune responses, disease mechanisms, and therapeutic strategies in physiologically relevant settings, with implications for both basic research and clinical practice in immunology and related fields. Here, we describe and discuss in detail a method to explore the in vivo elimination of target cells by antigen-specific CTLs using immunized/vaccinated mice as our experimental model.

Keywords: CD8 + T cell; CFSE, CTV; CTL; In vivo cytotoxic assay.

MeSH terms

  • Animals
  • Antigens* / immunology
  • Cytotoxicity, Immunologic*
  • Mice
  • T-Lymphocytes, Cytotoxic* / cytology
  • T-Lymphocytes, Cytotoxic* / immunology

Substances

  • Antigens