Nocardia opaca, a Gram-positive bacterium, is a potent source of immunostimulatory substances. Screening of sera of adult human donors revealed that all sera tested contained antibodies reactive with isolated Nocardia fractions (Nocardia delipidated cell mitogen, NDCM; Nocardia lysozyme digest, NLD; Nocardia water-soluble mitogen, NWSM; and fraction B). The respective values of reciprocal titres for IgM and IgG were in the range of 100 to 12,800, and 10 to 320 for IgA antibody isotypes, when NLD or fraction B were used as antigens in enzyme-linked immunosorbent assay (ELISA) tests. The level of antibodies directed to NDCM, a potent polyclonal B cell activator, was found to be the lowest. In vitro spontaneous as well as NDCM-induced production of antibodies to NDCM by human peripheral blood lymphocytes involved mainly the IgM class. Western-blot analysis demonstrated that antibodies in normal human sera react with nocardial antigens of molecular mass approximately 60, 40, 20 and 15-10 kDa. The same antigens were also recognized by rabbit and mouse hyperimmune sera, also confirming the immundominancy of these nocardial antigens in other species. The presence of anti-nocardia antibodies in human sera and their production by both stimulated and non-stimulated lymphocytes points to the natural sensitization of humans either by ubiquitous no-cardial components or by cross-reactive bacterial or food antigens.