In our previous study, a high rate of "heterophile antibody" was found in idiopathic cardiomyopathy. In the present investigation the cytotoxic activity of this antibody was examined in cultured myocardial cells. Trypsin treated cells cultured for 3 days were used in the two-step method of the complement dependent cytotoxicity test. The trypan blue dye-exclusion method was utilized to determine the percentage of surviving cells, and the cytotoxic effect was expressed by the cytotoxic index (CI). With fresh rabbit serum as complement, the mean CI value of the heterophile antibody positive sera was 20.7, which was in contrast to the value of anti-heart antibodies negative sera (p less than 0.01). As the cytotoxic effect to the heterophile antibody was absent without the complement, it may be complement dependent. Thus, our results suggest that the heterophile antibody may play a role in the pathogenesis or be an incremental factor of idiopathic cardiomyopathy.