We report the presence of a middle ear protein that has the same epitope as human surfactant protein A. Monoclonal antibodies (PC-6 and PE-10) against human pulmonary surfactant protein A stained faint granules of the mucosal epithelial cell cytoplasm in the orifice of the eustachian tube immunohistochemically. These antibodies also reacted with middle ear effusion of patients with otitis media with effusion by dot immunoassay, and recognized an 80 kd protein by Western blot analysis. These findings indicate that a protein immunoreactive with PC-6 and PE-10 occurs in the mucosal epithelial cells of the middle ear, and is also present in middle ear effusion. It is therefore likely that this 80 kd protein might be secreted from the mucosal epithelial cells to the middle ear cavity.