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Updated: Aug 3, 2025
In Vitro Tumor Cell Rechallenge For Predictive Evaluation of Chimeric Antigen Receptor T Cell Antitumor Function
Published on: February 27, 2019
Anti-tumor memory CD4 and CD8 T-cells quantified by bulk T-cell receptor (TCR) clonal analysis
Yanhua Gao1, Ira Bergman1,2,3
1Department of Pediatrics, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States.
Abstract:
Simple, reliable methods to detect anti-tumor memory T-cells are necessary to develop a clinical tumor vaccination program. A mouse model of curative viral onco-immunotherapy found that peritoneal tumor challenge following cure identified an oligoclonal anti-tumor memory CD4 and CD8 T-cell response. Clonotypes differed among the challenged animals but were congruent in blood, spleen and peritoneal cells (PC) of the same animal. Adoptive transfer demonstrated that the high-frequency responding T-cells were tumor specific. Tetramer analysis confirmed that clonotype frequency determined by T-cell receptor (TCR)- chain (TRB) analysis closely approximated cell clone frequency. The mean frequency of resting anti-tumor memory CD4 T-cells in unchallenged spleen was 0.028% and of memory CD8 T-cells was 0.11% which was not high enough to distinguish them from background. Stimulation produced a mean ~10-fold increase in splenic and 100-fold increase in peritoneal anti-tumor memory T-cell clonotypes. This methodology can be developed to use blood and tissue sampling to rapidly quantify the effectiveness of a tumor vaccine or any vaccine generating therapeutic T-cells.
Insights
Detecting anti-tumor memory T-cells is crucial for cancer vaccines. New methods using T-cell receptor analysis in mice show promise for quantifying vaccine effectiveness by measuring T-cell responses.
Area of Science:
- Immunology
- Oncology
- Vaccinology
Background:
- Developing effective clinical tumor vaccination programs requires reliable methods to detect anti-tumor memory T-cells.
- Viral onco-immunotherapy in a mouse model provided a system to study these responses.
Purpose of the Study:
- To establish and validate a method for detecting and quantifying anti-tumor memory T-cells.
- To assess the tumor specificity and frequency of these memory T-cells.
Main Methods:
- Utilized a mouse model of viral onco-immunotherapy.
- Induced peritoneal tumor challenge post-cure to identify memory T-cell responses.
- Employed T-cell receptor (TCR) repertoire analysis and tetramer staining.
- Performed adoptive transfer experiments to confirm T-cell specificity.
Main Results:
- Identified an oligoclonal anti-tumor memory CD4 and CD8 T-cell response upon tumor challenge.
- Demonstrated that T-cell clonotypes were congruent across blood, spleen, and peritoneal cells within individual animals.
- Confirmed high-frequency responding T-cells were tumor-specific.
- Showed that TCR TRB analysis accurately approximated T-cell clone frequency.
- Observed a significant increase in anti-tumor memory T-cell clonotypes after stimulation, particularly in peritoneal cells.
Conclusions:
- A reliable method for detecting anti-tumor memory T-cells was developed using TCR analysis.
- This methodology can quantify the effectiveness of tumor vaccines or other T-cell-generating vaccines.
- Blood and tissue sampling can be used to rapidly assess vaccine-induced therapeutic T-cell responses.
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