Multiplex IP-FCM reveals network-scale protein-protein interaction differences between pro-immunogenic and pro-tolerogenic TCR signaling (P1130)

Abstract

Abstract T cell receptor signaling involves interactions of various kinases, phosphatases, ubiquitin ligases and adaptor proteins. However, the network-scale protein-protein interactions (PPIs) that mediate signaling, as well as the key differences between pro-immunogenic (agonist) and pro-tolerogenic (antagonist) signals, remain unclear. We have developed a novel method to examine large networks of signaling PPIs, multiplex immunoprecipitation measured by flow cytometry (mIP-FCM). Using multiple classes of Luminex® microspheres, we created a 96-well-formatted matrix array capable of measuring 484 PPIs in a single overnight assay. The array is focused on the most proximal sign..