Protective monoclonal antibody defines a circumsporozoite-like glycoprotein exoantigen of Cryptosporidium parvum sporozoites and merozoites.
The apicomplexan protozoan parasite Cryptosporidium parvum causes a diarrheal disease in humans and other mammals for which specific therapy and immunoprophylaxis are unavailable. Passive immunization with Abs against whole C. parvum organisms has variable efficacy in immunocompromised or neonatal hosts. Because apical and surface-exposed zoite Ags of the Apicomplexa are critical to infectivity and targets of protective immunity, we examined the ability of mAbs generated against such Ags in C. parvum sporozoites to passively protect against infection and identify biologically relevant parasite molecules. A panel of mAbs was produced against affinity-purified native Ags using sporozoite apical- and surface-reactive mAb C4A1 as binding ligand. One resulting mAb, designated 3E2, elicited prominent morphologic changes in sporozoites and merozoites characterized by rapid and progressive formation, posterior movement, and release of membranous Ag-mAb precipitates. These changes had a striking resemblance to the malarial circumsporozoite precipitate (CSP) reaction. Sporozoite infectivity was completely neutralized after in vitro exposure to 3E2 and the CSP-like reaction. Furthermore, orally administered 3E2 completely prevented or markedly reduced infection in neonatal BALB/c mice. 3E2 bound to apical complex and surface molecules of zoites and was demonstrated in membranous precipitates by immunoelectron microscopy. In Western blots, 3E2 recognized multiple 46 to approximately 770 kDa sporozoite Ags and an approximately 1300-kDa Ag designated CSL, also expressed by merozoites. CSL was characterized as a soluble glycoprotein exoantigen released by infectious sporozoites. Further, CSL was determined to be the molecular species mechanistically involved in the CSP-like reaction by its identification in SDS-PAGE gels and Western blots of purified membranous precipitates. These findings indicate that CSL has a functional role in sporozoite infectivity and is a candidate molecular target for passive or active immunization against cryptosporidiosis.
Riggs
M W
MW
Department of Veterinary Science and Microbiology, University of Arizona, Tucson 85721, USA.
Stone
A L
AL
Yount
P A
PA
Langer
R C
RC
Arrowood
M J
MJ
Bentley
D L
DL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use*; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D002417:Cattle; D002846:Chromatography, Affinity; D003457:Cryptosporidiosis / Q000276:immunology* / Q000469:parasitology / Q000517:prevention & control*; D016785:Cryptosporidium parvum / Q000254:growth & development* / Q000276:immunology* / Q000648:ultrastructure; D005260:Female; D006801:Humans; D007116:Immunization, Passive; D016056:Immunodominant Epitopes / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D015800:Protozoan Proteins / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D016056:Immunodominant Epitopes; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan