Microbial adhesion of Cryptosporidium parvum sporozoites: purification of an inhibitory lipid from bovine mucosa.
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.
Johnson
Julie K
JK
Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802, USA.
Schmidt
Joann
J
Gelberg
Howard B
HB
Kuhlenschmidt
Mark S
MS
D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003457:Cryptosporidiosis / Q000517:prevention & control; D016785:Cryptosporidium parvum / Q000187:drug effects / Q000378:metabolism*; D006001:Glycoconjugates / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000737:chemistry / Q000378:metabolism / Q000469:parasitology*; D008055:Lipids / Q000302:isolation & purification / Q000494:pharmacology*; D008297:Male; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D009077:Mucins / Q000494:pharmacology
D006001:Glycoconjugates; D008055:Lipids; D009077:Mucins