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A synthetic cytotoxic T cell platform for rapidly prototyping TCR function.

Govinda Sharma¹, James Round¹, Fei Teng¹

¹Michael Smith Genome Sciences Centre, British Columbia Cancer Research Institute, Vancouver, BC, Canada.

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Summary

Researchers developed a new bioplatform for T cell receptor (TCR) profiling. This system enables rapid assessment of TCR cytotoxic potency and antigen recognition, overcoming limitations of current methods.

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Area of Science:

Immunology
Cell Biology
Biotechnology

Background:

Current methods for profiling T cell receptor (TCR) function, including cytotoxic potency and cross-reactivity, face challenges in establishing model systems for diverse HLA alleles and antigens.
Testing TCRs requires complex systems that can accommodate various HLA types and a wide range of potential antigens.

Purpose of the Study:

To develop a universal prototyping platform for functional profiling of T cell receptors (TCRs) with enhanced cytotoxic potency and cross-reactivity assessment.
To create a system enabling facile recombinant expression of TCR, peptide, and MHC-I coding sequences for direct response assessment.

Main Methods:

Implementation of a granzyme-activatable sensor for T cell cytotoxicity.
Engineering of an immortalized natural killer cell line (YT-Indy) and an MHC-null antigen-presenting cell line (K562) into a universal prototyping platform.
Integration of a fluorescence-based reporter for early detection of cytotoxic function and genetic interventions to preserve target cells.

Main Results:

Successful reconstitution of the surface TCR complex in the YT-Indy cell line at biologically relevant levels.
Demonstrated successful induction and highly sensitive detection of antigen-specific responses for multiple distinct model TCRs.
Confirmed preservation of target cells after 24-hour exposure to cytotoxic effectors in co-culture experiments.

Conclusions:

The developed bioplatform facilitates rapid expression and characterization of T cell receptor (TCR) responses.
This system empowers researchers to gain insights into TCR recognition patterns and optimize therapeutic TCRs.
The platform overcomes previous limitations in testing TCRs against diverse HLA alleles and antigens, advancing TCR-based research and therapeutics.