Methods of high-throughput identification of T cell epitopes by capturing cytokines on the surface of antigen-presenting cells

Abstract

The present disclosure relates to methods that combine identification of T cell-secreted cytokines by modified antigen-presenting cells (APCs), cell sorting, and next-generation sequencing to identify class I- and class II-restricted epitopes starting from massively-complex peptide-encoding oligonucleotide pools. APCs are modified to express anti-cytokine antibodies, a library of DNA-encoded peptides, and multiple HLA class I or II molecules. Upon co-culture with T cells, these modified APCs form HLA/epitope/TCR complexes, enabling the production of T cell activation-dependent cytokines with the DNA that encodes the presented peptide. After co-culture, the APCs are sorted, and the peptide-encoding DNA is sequenced to determine the identity of the immunogenic peptides. Thus, the disclosure allows pooled screening of thousands of encoded peptides to enable epitope discovery for orphan T cell receptors.

Dependencies are inferred from the claim text.