Profiling antigen-reactive T cells using high throughput single-cell T cell receptor sequencing.

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B942
Abstract ID: 5995

Presenting Author:
Efthymia Papalexi , Senior Scientist at Parse Biosci.

Abstract:

Recent advances in single-cell sequencing allow for simultaneous profiling of T cell receptors (TCRs) and transcriptomes leading to the characterization of key T cell subsets with antigen recognition and killing capabilities. Despite their success, these methods rely on microfluidic devices or plate-based protocols with limited sensitivity and throughput (1,000s-10,000s of cells, 1-2 samples) making the study of disease-relevant T cells time consuming and costly. 

To overcome these limitations, we developed a  split pool combinatorial barcoding technology to simultaneously characterize the TCRs alongside the full transcriptomes of up to 1 million T cells from up to 96 unique samples. Here, we profiled hundreds of thousands of human T cells stimulated with antigens from a variety of diseases. Applying our method, we performed deep characterization of antigen-reactive TCR clonotypes and assessed their sequence similarities. In addition, we performed transcriptome-based clustering analyses and identified highly proliferative and cytotoxic T cell subsets. Finally, we correlated T cell activation and cytotoxicity states to clonal expansion frequencies in order to identify TCRs with increased killing potential.

In summary, we present a highly flexible and scalable combinatorial barcoding technology to allow researchers to profile up to 1 million T cells in one experiment and investigate their functional responses during infection, cancer, autoimmunity, or therapeutic interventions.