ERM-Dependent Assembly of T Cell Receptor Signaling and Co-stimulatory Molecules on Microvilli prior to Activation

Abstract

Summary: T cell surfaces are covered with microvilli, actin-rich and flexible protrusions. We use super-resolution microscopy to show that ≥90% of T cell receptor (TCR) complex molecules TCRαβ and TCRζ, as well as the co-receptor CD4 (cluster of differentiation 4) and the co-stimulatory molecule CD2, reside on microvilli of resting human T cells. Furthermore, TCR proximal signaling molecules involved in the initial stages of the immune response, including the protein tyrosine kinase Lck (lymphocyte-specific protein tyrosine kinase) and the key adaptor LAT (linker for activation of T cells), are also enriched on microvilli. Notably, phosphorylated proteins of the ERM (ezrin, radixin, and moesin) family colocalize with TCRαβ as well as with actin filaments, implying a role for one or more ERMs in linking the TCR complex to the actin cytoskeleton within microvilli. Our results establish microvilli as key signaling hubs, in which the TCR complex and its proximal signaling molecules and adaptors are preassembled prior to activation in an ERM-dependent manner, facilitating initial antigen sensing. : T-cell surfaces are covered with microvilli, actin-rich and flexible protrusions. Ghosh et al. show that T cell receptor complex molecules, as well as several proximal signaling molecules, are preassembled on microvilli in an ERM-dependent manner. These results establish microvilli as key signaling hubs that facilitate initial antigen sensing by T cells. Keywords: T cell receptor, CD4, CD2, Lck, LAT, Ezrin-Radixin-Moesin (ERM) proteins, membrane protein clustering, actin cytoskeleton, total internal reflection microscopy, super-resolution microscopy