Characterization of the Cryptosporidium antigens from sporulated oocysts of Cryptosporidium parvum.
The antigenic constituents of sporulated Cryptosporidium parvum oocyst antigens were characterized with antisera from mice immunized against C. parvum. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining defined the major proteins. Six of seven lectins used recognized as many as 15 bands. The lectins concanavalin A, Dolichos biflorus, and wheat germ agglutinin showed strong activity against the same eight bands with molecular weights ranging from 72,000 to greater than 100,000. An enzyme-linked immunosorbent assay was used to detect antibody to C. parvum. Antibody binding was significantly decreased by heat and enzymatic treatment with trypsin, protease, and mixed glycosidases. C. parvum antigens were further defined by the reactivity of immune sera with a C. parvum sonicate preparation separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose paper. Antisera from orally infected mice consistently recognized four antigens with molecular weights ranging from 72,000 to greater than 100,000. These antigens also bound concanavalin A. Treatment of the antigen preparation with mixed glycosidases reduced the reactivity of antisera with most antigens with molecular weights greater than 60,000. The data suggest that the antigenic composition of C. parvum is complex and that carbohydrates alone or in association with lipids or proteins may be important in the immune response to C. parvum.
Luft
B J
BJ
Department of Medicine, State University of New York at Stony Brook.
Payne
D
D
Woodmansee
D
D
Kim
C W
CW
D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis*; D000953:Antigens, Protozoan / Q000032:analysis* / Q000276:immunology; D003044:Coccidia / Q000276:immunology*; D003457:Cryptosporidiosis / Q000276:immunology*; D003458:Cryptosporidium / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000032:analysis; D006026:Glycoside Hydrolases; D006358:Hot Temperature; D007106:Immune Sera / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D037102:Lectins; D051379:Mice; D009844:Oligosaccharides / Q000276:immunology; D010447:Peptide Hydrolases; D014357:Trypsin
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D006023:Glycoproteins; D007106:Immune Sera; D007074:Immunoglobulin G; D037102:Lectins; D009844:Oligosaccharides; D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases; D014357:Trypsin