Metalloendopeptidases from the intestinal brush border of Haemonchus contortus as protective antigens for sheep.
Substantial protection against the economically important nematode Haemonchus contortus has been achieved by immunizing sheep with a glycoprotein fraction isolated from the intestinal membranes of this parasite. This fraction has been termed Haemonchus galactose-containing glycoprotein complex (H-gal-GP) since it was originally isolated through its selective binding to lectins with a specificity for N-acetylgalactosamine. A major component of this highly protective antigen complex is a family of four zinc metalloendopeptidases, designated MEPs 1-4. Various combinations of these MEPs were evaluated in immunization-challenge trials in sheep. In two experiments a combination of all four MEPs, separated from the rest of the complex by gel filtration in 8 m urea, significantly reduced H. contortus egg counts by 45 and 50%, an effect not significantly different from that conferred by 8 m urea treatment of H-gal-GP itself. Similarly, MEP3 alone or MEPs 1, 2 and 4 in combination, electroeluted from the complex following SDS gel electrophoresis, each reduced egg counts by some 33%. The MEPs are therefore protective components of H-gal-GP and from previously published findings, it appears that MEP3 is the most effective member of this metalloendopeptidase family. However, there was no significant protection when sheep were immunized with fully reduced and denatured H-gal-GP or with bacterially expressed recombinant forms of MEP 1 or the principal domains of MEP3, suggesting that conformational epitopes on the MEPs are required for immunity.
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D002850:Chromatography, Gel / Q000662:veterinary; D010450:Endopeptidases / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005243:Feces / Q000469:parasitology; D005260:Female; D006188:Haemonchiasis / Q000201:enzymology / Q000276:immunology* / Q000469:parasitology / Q000517:prevention & control; D006189:Haemonchus / Q000201:enzymology / Q000276:immunology*; D015801:Helminth Proteins; D007114:Immunization / Q000662:veterinary; D015151:Immunoblotting; D007413:Intestinal Mucosa / Q000201:enzymology / Q000276:immunology*; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology*; D008666:Metalloendopeptidases / Q000276:immunology* / Q000302:isolation & purification; D008871:Microvilli / Q000201:enzymology / Q000276:immunology; D010270:Parasite Egg Count / Q000662:veterinary; D012756:Sheep; D012757:Sheep Diseases / Q000201:enzymology / Q000276:immunology / Q000469:parasitology*
D000909:Antibodies, Helminth; D015801:Helminth Proteins; D008562:Membrane Glycoproteins; D010450:Endopeptidases; D008666:Metalloendopeptidases; C119206:galactose-containing glycoprotein, Haemonchus contortus