Defining T cell receptor repertoires using nanovial-based affinity and functional screening.

Area of Science:

• Immunology
• Biotechnology
• Molecular Biology

Background:

• T cell receptor (TCR) immunotherapies hold therapeutic potential.
• Identifying antigen-reactive T cells and their specific TCRs is crucial for effective immunotherapy.
• Current methods for TCR identification and functional assessment can be limiting.

Approach:

• Utilized cavity-containing hydrogel microparticles (nanovials) coated with peptide-major histocompatibility complex (pMHC) monomers to capture and activate T cells.
• Enabled sorting of T cells based on both binding affinity and secreted cytokines.
• Employed microfluidic emulsion-based single-cell sequencing to recover paired TCR αβ-chains.
• Integrated oligonucleotide-barcoding of nanovials to accurately link TCR sequences to specific pMHC targets.

Key Points:

• Nanovials facilitate the isolation of antigen-reactive T cells by capturing them via pMHC interactions.
• Cytokine secretion by captured T cells allows for functional assessment and sorting.
• Oligonucleotide-barcoding ensures 100% accuracy in linking identified TCR sequences to their cognate antigens.
• This approach identified an expanded repertoire of functional TCRs targeting viral antigens with high specificity.

Conclusions:

• The nanovial platform provides a highly accurate and efficient method for isolating and characterizing functional T cells and their TCRs.
• This technology significantly advances the discovery of TCRs for therapeutic applications, particularly in viral antigen targeting.
• The developed method offers a superior alternative to standard techniques for TCR repertoire analysis in immunotherapy research.