title: Mediation of Cryptosporidium parvum infection in vitro by mucin-like glycoproteins defined by a neutralizing monoclonal antibody.
abstract: The protozoan parasite Cryptosporidium parvum is a significant cause of diarrheal disease worldwide. Attachment to and invasion of host intestinal epithelial cells by C. parvum sporozoites are crucial steps in the pathogenesis of cryptosporidiosis. The molecular basis of these initial interactions is unknown. In order to identify putative C. parvum adhesion- and invasion-specific proteins, we raised monoclonal antibodies (MAbs) to sporozoites and evaluated them for inhibition of attachment and invasion in vitro. Using this approach, we identified two glycoproteins recognized by 4E9, a MAb which neutralized C. parvum infection and inhibited sporozoite attachment to intestinal epithelial cells in vitro. 4E9 recognized a 40-kDa glycoprotein named gp40 and a second, >220-kDa protein which was identified as GP900, a previously described mucin-like glycoprotein. Glycoproteins recognized by 4E9 are localized to the surface and apical region of invasive stages and are shed in trails from the parasite during gliding motility. The epitope recognized by 4E9 contains alpha-N-acetylgalactosamine residues, which are present in a mucin-type O-glycosidic linkage. Lectins specific for these glycans bind to the surface and apical region of sporozoites and block attachment to host cells. The surface and apical localization of these glycoproteins and the neutralizing effect of the MAb and alpha-N-acetylgalactosamine-specific lectins strongly implicate these proteins and their glycotopes as playing a role in C. parvum-host cell interactions.
authors.affiliation: Division of Geographic Medicine and Infectious Diseases, Tupper Research Institute, New England Medical Center, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Cevallos
authors.forename: N
authors.initials: N
authors.lastname: Bhat
authors.forename: R
authors.initials: R
authors.lastname: Verdon
authors.forename: D H
authors.initials: DH
authors.lastname: Hamer
authors.forename: B
authors.initials: B
authors.lastname: Stein
authors.forename: S
authors.initials: S
authors.lastname: Tzipori
authors.forename: M E
authors.initials: ME
authors.lastname: Pereira
authors.forename: G T
authors.initials: GT
authors.lastname: Keusch
authors.forename: H D
authors.initials: HD
authors.lastname: Ward
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018938:Caco-2 Cells; D016785:Cryptosporidium parvum / Q000502:physiology*; D000939:Epitopes; D006031:Glycosylation; D006790:Host-Parasite Interactions; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009077:Mucins / Q000276:immunology / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D009077:Mucins