Thyroid peroxidase (TPO) evokes high-affinity, IgG-class autoantibodies [TPO autoantibodies (TPOAbs)] and TPO-specific T cells that are markers of thyroid infiltration or implicated in thyroid destruction, respectively. A diverse repertoire of human monoclonal TPOAbs, unparalleled in other autoimmune diseases, provides invaluable probes for investigating antibody epitopes. Human TPOAbs recognize an immunodominant region comprising overlapping A and B domains on conformationally intact TPO. Amino acids recognized by TPOAbs are located in the regions with homology to myeloperoxidase (MPO) and the complement control protein (CCP) but not in the epidermal growth factor (EGF)-like region. T cells recognize epitopes in the MPO-like region but not in the CCP- or EGF-like regions in humans. Monoclonal human TPOAbs modulate processing of TPO protein to provide peptides for some T cells. A human T cell clone expressed transgenically in mice induces lymphocytic infiltration and hypothyroidism. This T cell's epitope is only generated by thyrocyte processing of endogenous TPO. Further, intact TPO expressed in vivo is also required for induction of TPOAbs in mice that resemble human autoantibodies. Overall, some TPO-specific T cells and the majority of autoantibodies in humans develop in response to TPO presented by thyroid cells, rather than to TPO released by damaged thyrocytes.