We addressed the question of why naturally occurring, polyclonal thyroid peroxidase (TPO) autoantibodies have a restricted epitopic repertoire to an immunodominant region (IDR). We hypothesized that immunizing BALB/c mice with major histocompatibililty complex (MHC) class II compatible B lymphocytes (A20 cells) preferentially diverting TPO to the MHC class II pathway would produce TPO antibodies with an epitopic specificity similar to human autoantibodies, namely to the IDR. For this purpose we stably expressed in A20 cells a fusion protein of TPO sandwiched between the signal peptide and transmembrane/cytoplasmic tail of the lysosome- associated membrane protein (LAMP) 1. Expression of LAMP1-TPO in A20 B cells was confirmed by flow cytometry using a TPO monoclonal antibody. Mice injected intraperitoneally with LAMP1-TPO A20 B cells developed TPO antibodies detected by enzyme-linked immunosorbent assay (ELISA), flow cytometry and (125)I-TPO precipitation. However, TPO antibody levels were low. Most important, competition for TPO antibody binding by recombinant human TPO autoantibody Fab indicated that more than 80% of the polyclonal TPO antibodies in the immunized mice were to the human autoantibody IDR. In summary, injecting mice with B lymphocytes that constitutively express TPO diverted to the MHC class II pathway generates antibodies with epitopes similar to those of human TPO autoantibodies, namely to the autoantibody IDR. However, these antibodies are of low titer that is itself associated with this epitopic bias.