Characterization of anti-Neospora caninum hyperimmune rabbit serum by western blot analysis and immunoelectron microscopy

Abstract

The antigens recognized by hyperimmune rabbit serum raised against tachyzoites of the NC-1 isolate of Neospora caninum were characterized using chemiluminescent Western blotting, immunogold-silver staining and immunoelectron microscopy. Approximately 20 immunodominant antigens whose relative rates of migration were 16-80 kDa were recognized by the serum in Western blots using reduced or nonreduced parasite antigen preparations. The nonreduced parasite antigens were more strongly recognized by the serum than were the reduced antigen preparations. Immunoelectron microscopy revealed that the rabbit-serum-labeled antigens were localized to some organelles of N. caninum tachyzoites and to the parasitophorous vacuole surrounding them. In particular, antigens found in the dense granules, in the micronemes, and in the posterior portion of the rhoptries were strongly labeled by an indirect immunogold-labeling technique.