Microbial adhesion of Cryptosporidium parvum sporozoites: Purification of an inhibitory lipid from bovine mucosa

Julie K. Johnson, Joann Schmidt, Howard B. Gelberg, Mark S. Kuhlenschmidt

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Cryptosporidium parvum is a protozoan pathogen of humans and livestock worldwide. Its ability to infect a wide range of species raises questions as to the involvement of a specific host cell receptor for parasite-host recognition. To investigate the mechanism of parasite-host cell recognition, we have developed an in vitro cell suspension binding assay to investigate adhesion of C. parvum sporozoites to host cells. Morphologic features of binding events observed with this assay were identical to those described in natural infections. Glycoconjugates, Madin Darby bovine kidney (MDBK) cell fractions, and plasma membrane vesicles (PMVs) were screened for their ability to block binding of sporozoites to MDBK cells. Mucins, MDBK cell fractions, and PMVs exhibited dose-dependent inhibition of sporozoite binding. The major inhibitory fraction from MDBK cells was found to be insoluble in aqueous medium, nonsaponifiable, and lacking carbohydrate moieties, nitrogen, and phosphorus. Its inhibitory effect was resistant to heat, protease digestion, and glycosidase treatment, suggesting that the inhibitory activity is a lipid or a lipid-like component. The inhibitory activity was purified from MDBK cells, and in larger amounts from bovine small intestinal mucosa, by organic solvent extraction, semipreparative high-pressure liquid chromatography, and preparative high-performance thin-layer chromatography. Biochemical analyses, thin-layer chromatography staining techniques, mass spectrometry, and elemental analysis were used to partially characterize the purified lipid. These results indicate that a host intestinal lipid(s) or a lipid-like component(s) may play an important role in the early stages of host cell invasion by C. parvum.

Original languageEnglish (US)
Pages (from-to)980-990
Number of pages11
JournalJournal of Parasitology
Issue number5
StatePublished - Oct 2004

ASJC Scopus subject areas

  • Parasitology
  • Ecology, Evolution, Behavior and Systematics


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