Abstract

The precise and efficient detection of epitope-specific T-cell responses is crucial for understanding adaptive immunity in health and disease, particularly in the context of cancer, autoimmunity, and infectious diseases. Current methods often suffer from low throughput or require complex, time-consuming cell manipulation. Here, we propose a novel, high-throughput approach for the **multiplexed detection of epitope-specific T-cell responses** that exploits the natural phenomenon of **trogocytosis**. Trogocytosis, the active transfer of membrane fragments and associated proteins between cells, is a key mechanism in immune cell communication. Our method leverages the transfer of the B-cell marker **CD20** from target cells to activated T-cells upon specific antigen recognition and immunological synapse formation. By differentially labeling target cells with distinct epitope-MHC complexes, we can simultaneously present multiple antigens in a single reaction volume. Flow cytometric analysis then allows for the rapid and quantitative identification of T-cells that have acquired the CD20 marker, serving as a direct readout of antigen-specific activation. This technique significantly enhances the speed and complexity of T-cell monitoring, offering a powerful new tool for high-content screening and personalized immunomonitoring with broad applications in membrane and cell biology research.