Angiostrongylus cantonensis: immunoblot analysis of the antigens recognized by rats

Following infection of rats with Angiostrongylus cantonensis, occurrence of anti-parasite antibody in the serum was determined with special reference to the antigens recognized by host IgG antibodies, using SDS-PAGE combined with an immunoblotting technique. Three saline extracts of digestive organ, reproductive organ and body wall, isolated from adult female A. cantonensis, were used as crude antigenic solutions. Then 7 to 49 days after infection, IgG antibodies directed predominantly against a single antigen, referred to as Ac-1 antigen, were detected. After 91 days or more, infected rats formed antibodies not only against the Ac-1 antigen, but also against a wide variety of other components with molecular weights in the range of 26,000-220,000 dalton. By using an antiserum against Ac-1 antigen as a probe, it was shown that the molecular weight and subunit structure, as well as the immunoelectrophoretic mobility, varied according to the organ from which the antigenic extract was prepared. The Ac-1 protein in the extract of the reproductive organ, one of the major sources of the Ac-1 antigen, showed the same electrophoretic mobility as alpha-globulin. It has a molecular weight in the range of 100,000-200,000 dalton under both non-reducing and reducing conditions. Immunohistochemical studies, using the same antiserum and sectioned adult female worms, found Ac-1 antigen in the cytoplasm of oocytes at different stages of development, and in the lateral cord.