T-cell Cytotoxicity Assays (CD8+ T-cell killing assay)

T-cell cytotoxicity assays, particularly CD8+ T-cell killing assays, are essential tools for validating neoantigens by measuring the ability of these T cells to kill target cells expressing the specific neoantigen. These assays help assess the immunogenicity of neoantigens and their potential effectiveness in cancer immunotherapy.

← Assays

Principle of the T-cell Cytotoxicity Assay

The CD8+ T-cell killing assay measures the ability of CD8+ T cells to recognize and kill target cells presenting specific neoantigens through MHC class I molecules. Upon recognition, T cells become activated and release cytotoxic factors, such as perforin and granzymes, that induce apoptosis in the target cells. The assay quantifies the extent of target cell lysis to assess the immunogenicity of neoantigens and the effectiveness of T-cell responses.

Model Description

Target cells expressing the neoantigen are co-cultured with effector cells (CD8+T cells), resulting in target cell destruction.

Readouts

Tumor cell destruction by flow cytometry and Effector cell response by cytokine secretion (IFNγ release)

Add-On Services

Evaluation of cytokine release, proteomics, and transcriptomics.

Key Components of CD8+ T-cell killing assay

A. Target Cells (Neoantigen-Expressing Cells)

  • Carboxyfluorescein succinimidyl ester (CFSE)-labeled target cells can be tumor cell lines or engineered cell lines that express the specific neoantigen of interest.

B. Effector cells (CD8+ T Cells)

C. Effector-to-Target (E:T) Ratios

  • Various E:T ratios (e.g., 1:1, 5:1, 10:1) will be tested to evaluate the cytotoxic potential of CD8+ T cells against target cells.

C. Detection Method

  • The extent of target cell killing is calculated based on the percentage of viable target cells remaining after co-culture with CD8+ T cells.