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Updated: Jun 13, 2026
Analysis of Human T Cell Activity in an Allogeneic Co-Culture Setting of Pre-Treated Tumor Cells
Published on: March 7, 2025
T cell activation upon exposure to patient-derived tumor tissue: a functional assay to select patients for adoptive T
Caroline Schroten1, Robert Kraaij, Joke L M Veldhoven
1Dept Urology, Erasmus MC, Rotterdam, The Netherlands.
Abstract:
Gene-engineered T cell therapy represents a promising strategy to treat cancers. To enable pre-selection of patients sensitive to this type of treatment we have setup and validated a T cell activation assay to test antigen expression on patient-derived tumor tissues. Chimeric antibody-based receptor (CAR) directed against CAIX, currently used in a clinical trial to treat RCC patients, was used as a model receptor. Primary human T cells expressing CAIX CAR were able to respond to CAIX-positive but not CAIX-negative tumor tissue and showed an increased production of IFNgamma, TNFalpha, IL-10 and IL-4, but not IL-2 or IL-5. Tumor tissue driven responses of primary T cells were paralleled by NFAT activation measured in CAR-transduced Jurkat T cells, which was shown to be triggered in a CAR and antigen-specific manner. Next, the reporter gene assay was applied to two independent PSMA CARs, which both mediated NFAT activation in response to tumor tissue. Taken together, a sensitive and donor-independent assay was established to measure T cell activation upon exposure to patient-derived tumor tissue, which may facilitate pre-selection of patients for clinical adoptive T cell therapy.
Insights
A new T cell activation assay identifies patients likely to benefit from cancer therapies. This assay measures T cell responses to tumor tissues, aiding in the selection of suitable candidates for adoptive T cell therapy.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Gene-engineered T cell therapy shows promise for cancer treatment.
- Patient selection is crucial for optimizing the efficacy of T cell therapies.
- Current methods lack efficient ways to pre-assess patient sensitivity.
Purpose of the Study:
- To develop and validate a T cell activation assay for pre-selecting cancer patients.
- To assess antigen expression on patient-derived tumor tissues.
- To enable patient stratification for adoptive T cell therapy.
Main Methods:
- Established a T cell activation assay using patient-derived tumor tissues.
- Utilized chimeric antigen receptor (CAR) T cells targeting CAIX and PSMA.
- Measured cytokine production (IFNγ, TNFα, IL-10, IL-4) and NFAT activation.
Main Results:
- CAIX CAR T cells responded specifically to CAIX-positive tumor tissue.
- T cell activation correlated with increased IFNγ, TNFα, IL-10, and IL-4 production.
- NFAT activation in reporter cells mirrored T cell responses in a CAR and antigen-specific manner.
- The assay successfully detected responses mediated by two independent PSMA CARs.
Conclusions:
- A sensitive, donor-independent assay for measuring T cell activation against tumor tissue was established.
- This assay can facilitate patient pre-selection for clinical adoptive T cell therapy.
- The findings support the use of this assay to improve treatment outcomes in cancer immunotherapy.
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