A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies

Abstract

Two hybridoma clones, CMYL3 and CMYL30, were generated by immunizing Balb/c mice with excysted oocysts of Cryptosporidium muris. Both clones secreted monoclonal antibodies against an oocyst-wall antigen with apparent molecular mass of 250 kDa (called CM250) from C. muris and C. parvum. The epitope appeared to be periodate-sensitive, suggesting the involvement of a carbohydrate moiety. Immunofluorescence and confocal microscopy on purified oocysts and infected mouse tissues revealed staining confined to the oocyst wall of both Cryptosporidium species. Immunogold labeling further revealed the presence of the CM250 antigen in electron-dense vesicles and cytoplasm of developing macrogametocytes, and ultimately localized to the oocyst wall of mature oocysts. Both antibodies cross-reacted with C. serpentis oocysts but did not recognize the other enteropathogenic protozoans Giardia muris, Eimeria falciformis and E. nischulz. These antibodies may be valuable tools for the analysis of oocyst-wall formation in Cryptosporidium and characterization of the common antigen.