Rat and human colonic mucins bind to and inhibit adherence lectin of Entamoeba histolytica.
Establishment of adherence by Entamoeba histolytica is mediated by a 170-kD Gal/GalNAc inhibitable lectin and is required for cytolysis and phagocytosis of mammalian target cells. We studied the biochemical mechanisms of the in vitro interaction between rat and human colonic mucins and axenic E. histolytica trophozoites. Crude mucus prevented amebic adherence to Chinese hamster ovary (CHO) cells by up to 70%. Purification of the colonic mucins by Sepharose 4B chromatography, nuclease digestion, and cesium chloride gradient centrifugation resulted in a 1,000-fold enrichment of the inhibitory mucins. Purified rat mucin inhibited amebic adherence to and cytolysis of homologous rat colonic epithelial cells. Oxidation and enzymatic cleavage of rat mucin Gal and GalNAc residues completely abrogated mucin inhibition of amebic adherence. The binding of rat 125I-mucin to amebae was galactose specific, saturable, reversible, and pH dependent. A monoclonal antibody specific for the 170-kD amebic Gal/GalNAc lectin completely inhibited the binding of rat 125I-mucin. Rat mucin bound to Affigel affinity purified the amebic lectin from conditioned medium. Colonic mucin glycoproteins act as an important host defense by binding to the parasite's adherence lectin, thus preventing amebic attachment to and cytolysis of host epithelial cells.
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002846:Chromatography, Affinity; D003106:Colon / Q000032:analysis*; D006224:Cricetinae; D004748:Entamoeba histolytica / Q000502:physiology*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D009077:Mucins / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*; D010084:Oxidation-Reduction; D051381:Rats; D011919:Rats, Inbred Strains
D037102:Lectins; D009077:Mucins; D006026:Glycoside Hydrolases