Hyperacute rejection of vascularized, discordant xenografts is generally though to be initiated when natural antibodies of the recipient bind to endothelial cells of the donor organ. While rejection of such xenografts always occurs, the molecular targets of natural antibodies have not been elucidated. The aim of the experiments reported herein was to identify the molecules on porcine endothelial cells that would be recognized by human natural antibodies if a porcine organ were to be transplanted into a human (or rhesus). Toward the end, it was shown that the major components recognized by human serum on porcine endothelial cells are glycoproteins of 115kDa, 125kDa, and 135kDa (gp115/135). Reactivity with these glycoproteins was abrogated by enzymatic cleavage of N-linked oligosaccharides or of subterminal beta-D-gal residues suggesting that the determinants are located on oligosaccharides rather than on the polypeptide cores. The biological relevance of gp115/135 was suggested by experiments in which a similar series of components was shown to be recognized by rhesus natural antibodies and by the absorption of such antibodies by perfusion of porcine kidneys. The gp115/135 antigens were present on porcine platelets but not porcine RBC or lymphocytes. Nevertheless, purified RBC and lymphocytes absorbed human anti-gp115/135, suggesting that human natural antibodies recognize the same or crossreactive carbohydrate determinants expressed on the surface of a variety of cells.