Multiplexed Detection of Epitope Specific T-Cell Responses Using CD20 Trogocytosis

Discovery of antigen epitopes and cognate T-cell receptors (TCRs) is one of the central challenges in the development of targeted immunotherapies. In vitro testing for antigen reactivity is the major protocol for these purposes. Here we have proposed an approach for multiplexing of the antigens/epitopes in one test volume with the unmixing of stimulus during the analysis. We found that B-cell marker CD20 is transferred from B-cells to T-cells surface during antigen-specific interactions. We demonstrated that anti-CD20 antibodies with different labels can be used as tracers of the specific antigens loaded into the B-cells, so that T-cells stimulated by certain antigen accept corresponding anti-CD20 antibodies. This approach can be expanded using the mixture of anti-CD20 antibodies that will increase the output and reliability of antigen discovery routine.