ABSTRACT
In this study, monoclonal antibodies were developed from the partially purified surface tegumental antigens of F. gigantica. Nine MoAbs: 2G11, 1G2, 1B12, 2G2, 2G5, 3C6, 3G2, 3G3 and 3F6 were used for anatomical localization of adult F. gigantica. The reaction was demonstrated by Avidin-Biotin method. The results revealed that among the sections stained with non-immune sera and control group, there were no reaction products on either the tegument or the cecal epithelium. The only brownish areas were the vitelline glands. In the sections stained with immune sera, brownish reaction products appeared on the surface membrane, the spine membrane, the cecal lumen and its epithelial cells. The experiment sections of nine monoclonal antibodies revealed that the reaction occurred mainly on the tegument of the adult worm which covered its surface and spine.
Subject(s)
Animals , Antibodies, Monoclonal , Antigens, Helminth/analysis , Fasciola/immunology , Fascioliasis/diagnosis , Immunoenzyme TechniquesABSTRACT
Monoclonal antibodies (MAbs) were produced by the in vitro fusion of Balb/C mice spleen cells immunized with partially purified surface tegument antigens of Fasciola gigantica. The surface membrane and tegument antigens were purified by using gel filtration chromatography. SDS-PAGE performed on the processed proteins demonstrated that the proteins had molecular weights of 20 to 97 kDa. In this study, fifteen monoclonal antibody clones were selected from the hybridoma clones, namely: 1B7, 1B11, 1B12, 1C9, 1D4, 1G2, 1H7, 2B6, 2C3, 2C9, 2D11, 2F11, 2G2, 2G5, and 2G11. They were evaluated by immunoblot assay and were differentiated into two groups. In the first group were found 60 and 38 kDa proteins; in the second group were found 66, 60, and 38 kDa proteins. All were found to secrete IgM, kappa light-chain antibodies. These MAbs were tested for their cross-reactivity with other trematodes commonly found to infect cattle and man. All of these MAbs showed some degree of cross-reactivity with other trematode species.