Identification and Characterization of Human Autoreactive T Cells for Autoimmune Research

T cells directed against self-antigens play a pivotal role in autoimmunity, yet their comprehensive characterization in human autoimmune disorders remains challenging. In this chapter, we present methods to isolate and profile autoreactive T cells from human biological samples, specifically peripheral blood, cerebrospinal fluid, and tissue biopsies. The workflow combines in vitro T cell screening, isolation of autoreactive T cells, and single T cell clone generation, followed by in depth analysis of T cell receptor (TCR) repertoire, HLA restriction, cytokine profile, and targeted self-epitope. These protocols aim to facilitate research of human autoimmune diseases by providing powerful tools for systematic analysis of autoreactive T cells in human biological samples.