Related Experiment Video
Updated: Nov 4, 2025
07:05
In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model
Published on: November 28, 2017
10.1K
Antigen-Specific In Vivo Killing Assay
Nada Chaoul1, Gilles Dadaglio2
1Department of Emergency and Organ Transplant, School and Chair of Allergology and Clinical Immunology, University of Bari-Aldo Moro, Bari, Italy. nada.chaoul@uniba.it.
Methods in Molecular Biology (Clifton, N.J.)
|May 30, 2021
Summary
This study details an in vivo killing assay to measure the antigen-specific killing capacity of Cytotoxic CD8+ T Lymphocytes (CTLs). The protocol quantifies CTL lysis of cells presenting the SIINFEKL peptide from the OVA protein.
Area of Science:
- Immunology
- Cellular immunology
- T cell biology
Background:
- Cytotoxic CD8+ T Lymphocytes (CTLs) are crucial for adaptive immunity.
- CTLs recognize and eliminate target cells presenting foreign or modified peptides via MHC class I molecules.
- Quantifying antigen-specific CTL activity in vivo is essential for understanding immune responses.
Purpose of the Study:
- To describe a detailed protocol for an in vivo killing assay.
- To quantify the antigen-specific killing capacity of CTLs.
- To demonstrate the lysis of target cells presenting the H-2 Kb-restricted OVA257-264 (SIINFEKL) epitope.
Main Methods:
- Development and validation of an in vivo CTL killing assay.
- Use of target cells engineered to express the H-2 Kb-restricted OVA257-264 (SIINFEKL) epitope.
- Quantification of specific cell lysis in vivo.
Main Results:
- The assay effectively quantifies antigen-specific CTL killing capacity in mice.
- Demonstrated lysis of target cells presenting the SIINFEKL epitope.
- Established a reproducible method for assessing CTL function.
Conclusions:
- The described in vivo killing assay is a valuable tool for studying CTL-mediated immunity.
- This protocol facilitates the assessment of immune responses against specific antigens.
- Provides a foundation for further research in T cell immunology and immunotherapy.
