Chagas disease cardiomyopathy (CCC) is one of the few examples of post-infectious autoimmunity, where infectious episodes with an established pathogen, the protozoan parasite Trypanosoma cruzi, clearly triggers molecular mimicry-related target organ immune damage. CD4+ T-cell clones infiltrating hearts from CCC patients cross-reactively recognize human cardiac myosin, the major heart protein, and the immunodominant B13 protein from T. cruzi. Moreover, in vitro priming with B13 leads to the recovery of cardiac myosin cross-reactive T-cell clones. In order to identify cross-reactive epitopes between B13 protein and human cardiac myosin, we used B13 peptide S15.4, preferentially recognized by CCC patients, to establish a T-cell clone from an HLA-DQ7 individual. The B13 S15.4 peptide-specific CD4+ T-cell clone 3E5 was tested in proliferation assays against 15 Lys/His-substituted S15.4-derived peptides for TCR/HLA contact analysis. Together with previous HLA-binding data and molecular modeling of the HLA-DQ7-peptide S15.4 complex, Lys/His scanning analysis showed eight TCR/HLA contact positions. Clone 3E5 was also tested against 45 15-mer peptides from human beta-cardiac myosin heavy chain bearing the central HLA-DQ7 binding motif. Clone 3E5 recognized 13 peptides from cardiac myosin. The alignment of cross-reactive peptides in cardiac myosin showed very limited sharing of residues or side chains with similar chemical/structural features at aligned positions, indicative of a very degenerate TCR recognition pattern. The existence of degenerate intramolecular recognition, with multiple low-homology, cross-reactive epitopes in a single autoantigenic protein may have implications in increasing the magnitude of the autoimmune response in CCC and other autoimmune diseases.