We determined the response of peripheral blood mononuclear cells from patients with bullous pemphigoid and normal subjects to synthetic peptides encoded by BPAG1. Peripheral blood mononuclear cells from patients and normal subjects were cocultured in the presence of 15-22-amino-acid-long amphipathic and hydrophilic peptides selected from the BPAG1 sequence. Seven of 10 patients (70%) with bullous pemphigoid had an increased response of peripheral blood mononuclear cells (> 3.25/10(6) cells) when cultured with amphipathic sequences encoded by BPAG1 compared to 3 of 10 (30%) normal subjects. Peripheral blood mononuclear cells from 3 of 15 (20%) patients and 3 of 15 normal subjects (20%) demonstrated an increased response when cultured with hydrophilic peptides. Peptides associated with an increased peripheral blood mononuclear cell response in patients with bullous pemphigoid were adjacent to regions of BPAG1 recently demonstrated to contain epitopes recognized by circulating autoantibodies in the sera of patients with bullous pemphigoid. Increased peripheral blood mononuclear cell responses were more commonly observed in patients with bullous pemphigoid with generalized disease and those who had their disease for longer than 2 months (p < 0.05). The observation that increased duration and generalized disease was associated with increased peripheral blood mononuclear cell responses to peptides encoded by BPAG1 supports the hypothesis that responses to BPAG1 may occur as a consequence of ongoing inflammation at the basement membrane.