title: The Cryptosporidium parvum C-Type Lectin CpClec Mediates Infection of Intestinal Epithelial Cells via Interactions with Sulfated Proteoglycans.
abstract: The apicomplexan parasite Cryptosporidium causes significant diarrheal disease worldwide. Effective anticryptosporidial agents are lacking, in part because the molecular mechanisms underlying Cryptosporidium-host cell interactions are poorly understood. Previously, we identified and characterized a novel Cryptosporidium parvum C-type lectin domain-containing mucin-like glycoprotein, CpClec. In this study, we evaluated the mechanisms underlying interactions of CpClec with intestinal epithelial cells by using an Fc-tagged recombinant protein. CpClec-Fc displayed Ca(2+)-dependent, saturable binding to HCT-8 and Caco-2 cells and competitively inhibited C. parvum attachment to and infection of HCT-8 cells. Binding of CpClec-Fc was specifically inhibited by sulfated glycosaminoglycans, particularly heparin and heparan sulfate. Binding was reduced after the removal of heparan sulfate and following the inhibition of glycosaminoglycan synthesis or sulfation in HCT-8 cells. Like CpClec-Fc binding, C. parvum attachment to and infection of HCT-8 cells were inhibited by glycosaminoglycans and were reduced after heparan sulfate removal or inhibition of glycosaminoglycan synthesis or sulfation. Lastly, CpClec-Fc binding and C. parvum sporozoite attachment were significantly decreased in CHO cell mutants defective in glycosaminoglycan synthesis. Together, these results indicate that CpClec is a novel C-type lectin that mediates C. parvum attachment and infection via Ca(2+)-dependent binding to sulfated proteoglycans on intestinal epithelial cells.
authors.affiliation: Tufts University Sackler School of Graduate Biomedical Sciences, Boston, Massachusetts, USA.
authors.forename: Jacob G
authors.initials: JG
authors.lastname: Ludington
authors.affiliation: Tufts University Sackler School of Graduate Biomedical Sciences, Boston, Massachusetts, USA Hward@Tuftsmedicalcenter.org.|Division of Geographic Medicine and Infectious Disease, Tufts Medical Center, Boston, Massachusetts, USA.
authors.forename: Honorine D
authors.initials: HD
authors.lastname: Ward
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D016785:Cryptosporidium parvum / Q000502:physiology*; D004705:Endocytosis; D004847:Epithelial Cells / Q000469:parasitology*; D054884:Host-Pathogen Interactions; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism*; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism*
chemical_list: D037181:Lectins, C-Type; D011509:Proteoglycans