title: An AC-5 cathepsin B-like protease purified from Haemonchus contortus excretory secretory products shows protective antigen potential for lambs.
abstract: The immunogenic properties of cysteine proteases obtained from excretory/secretory products (ES) of Haemonchus contortus were investigated with a fraction purified with a recombinant H. contortus cystatin affinity column. The enrichment of H. contortus ES for cysteine protease was confirmed with substrate SDS-PAGE gels since the cystatin-binding fraction activity was three times higher than total ES, despite representing only 3% of total ES. This activity was inhibited by a specific cysteine protease inhibitor (E64) and by recombinant cystatin. The one-dimensional profile of the cystatin-binding fraction displayed a single band with a molecular mass of 43 kDa. Mass spectrometry showed this to be AC-5, a cathepsin B-like cysteine protease which had not been identified in ES products of H. contortus before. The cystatin binding fraction was tested as an immunogen in lambs which were vaccinated three times (week 0, 2.5 and 5), challenged with 10 000 L3 H. contortus (week 6) before necropsy and compared to unvaccinated challenge controls and another group given total ES (n = 10 per group). The group vaccinated with cystatin-binding proteins showed 36% and 32% mean worm burden and eggs per gram of faeces (EPG) reductions, respectively, compared to the controls but total ES was almost without effect. After challenge the cystatin-binding proteins induced significantly higher local and systemic ES specific IgA and IgG responses.
authors.affiliation: Division of Parasitology and Tropical Veterinary Medicine, Department of Infectious Diseases and Immunology, Utrecht University, Utrecht, The Netherlands.
authors.forename: Erik
authors.initials: E
authors.lastname: De Vries
authors.forename: Nicole
authors.initials: N
authors.lastname: Bakker
authors.forename: Jeroen
authors.initials: J
authors.lastname: Krijgsveld
authors.forename: Dave P
authors.initials: DP
authors.lastname: Knox
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.forename: Ana Patricia
authors.initials: AP
authors.lastname: Yatsuda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology*; D049109:Cell Proliferation; D002846:Chromatography, Affinity; D003546:Cysteine Endopeptidases / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006188:Haemonchiasis / Q000276:immunology / Q000662:veterinary*; D006189:Haemonchus / Q000201:enzymology*; D008214:Lymphocytes / Q000378:metabolism; D011485:Protein Binding; D012756:Sheep; D012757:Sheep Diseases / Q000469:parasitology / Q000517:prevention & control*; D013997:Time Factors; D014612:Vaccines / Q000276:immunology*
chemical_list: D000947:Antigens, Helminth; D014612:Vaccines; D003546:Cysteine Endopeptidases