Generation of monoclonal antibodies to characterize antigens of the nematode Nippostrongylus brasiliensis.

Hybridomas secreting monoclonal antibodies to Nippostrongylus brasiliensis antigens were generated by fusion of NSI mouse myeloma cells with spleen cells from Balb/c mice which had been immunized either with killed third stage larvae or killed adult worms of N. brasiliensis. Enzyme-linked immunosorbent assays with worm or larvae fragments as antigens revealed that thirteen hybrids produced parasite-specific antibodies. The antibodies reacted with either both worms and larvae or exclusively recognized worm antigens. No antibody was found which solely reacted with larval antigens. Immunofluorescence studies demonstrated that most antibodies homogeneously stained the cuticle of 80-90% of the adult worms and 10-20% of the third stage larvae. The antibodies were further characterized by enzyme-linked immunosorbent assays specific for worm or larvae homogenates and secretions or for acetylcholinesterase from electric eel and bovine erythrocytes. Investigations of the epitope specificity by the inhibition of binding to worm fragments in the presence of various glycosides, N-acetylneuraminic acid and phosphorylcholine revealed that six antibodies reacted with N-acetylneuraminic acid. All antibodies displayed a low affinity to phosphorylcholine.