High-Throughput Screening of Functional Neo-Antigens and Their Specific T-Cell Receptors via the Jurkat Reporter System Combined with Droplet Microfluidics

Abstract

T-cell receptor (TCR)-engineered T cells can precisely recognize a broad repertoire of targets derived from both intracellular and surface proteins of tumor cells. TCR-T adoptive cell therapy has shown safety and promising efficacy in solid tumor immunotherapy. However, antigen-specific functional TCR screening is time-consuming and expensive, which limits its application clinically. Here, we developed a novel integrated antigen–TCR screening platform based on droplet microfluidic technology, enabling high-throughput peptide–major histocompatibility complex (pMHC)-to-TCR paired screening with a high sensitivity and low background signal. We introduced DNA barcoding technology to label peptide antigen candidate-loaded antigen-presenting cells and Jurkat reporter cells to check the specificity of pMHC–TCR candidates. Coupled with the next-generation sequencing pipeline, interpretation of the DNA barcodes and the gene expression level of the Jurkat T-cell activation pathway provided a clear peptide–MHC–TCR recognition relationship. Our proof-of-principle study demonstrates that the platform could achieve pMHC–TCR paired high-throughput screening, which is expected to be used in the cross-reactivity and off-target high-throughput paired testing of candidate pMHC–TCRs in clinical applications.

Editorial: Quantification and prediction of T-cell cross-reactivity through experimental and computational methods

“…Unfortunately, the complexity and diversity of the molecules involved in these interactions has slowed the development of both experimental and computational methods to detect, quantify, and predict these T-cell cross-reactivity events. But this picture has been changing in recent years, with exciting developments in both high-throughput experimental methods ( 5 , 14 , 15 ) and scalable computational approaches ( 16 – 21 ) for the study of TCRpMHC interactions. This Research Topic highlights recent studies in this field, including further analysis of T-cell cross-reactivity in antiviral immunity, and new computational approaches to predict TCR specificity and off-target toxicity.…”