Haemonchus contortus: characterization of the baculovirus expressed form of aminopeptidase H11.
Recombinant form of Haemonchus contortus aminopeptidase H11, an intestinal membrane glycoprotein considered to be in its native form the most promising vaccine candidate, was produced in insect cells, characterised and tested in pilot vaccination-challenge trial on sheep. The sequence of the cloned gene, obtained by RT PCR isolated from adult worms, showed 97% identity to the highly immunogenic H11 clone, described by Graham et al., (database accession number AJ249941.1). A 1305 bp fragment of H11 was expressed in E. coli and used to raise a specific antiserum, which recognized recombinant forms of H11 and 110 kDa protein from H. contortus extract. H11 was expressed by baculovirus recombinants in insect cells in full length and as a fusion protein with H. contortus glutathione S-transferase (GST). The baculovirus produced recombinant antigens were used without adjuvants to immunize sheep, which resulted in 30% (full length H11) and 20% (GST-H11) reduction of worm burden. These animal experiments indicated that, although the protection induced by in vitro produced protein is lower than in case of H11 isolated from worms, recombinant forms of aminopeptidase may be considered as antigens for the control of haemonchosis.
Reszka
Natalia
N
University of Gdańsk, Department of Molecular Virology, Kładki 24, 80-822 Gdańsk, Poland.
Rijsewijk
Frans A M
FA
Zelnik
Vladimir
V
Moskwa
Bozena
B
Bieńkowska-Szewczyk
Krystyna
K
D000626:Aminopeptidases / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D016367:Baculoviridae; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D010450:Endopeptidases / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006188:Haemonchiasis / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D006189:Haemonchus / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D007114:Immunization / Q000379:methods / Q000662:veterinary; D007313:Insecta; D012333:RNA, Messenger / Q000302:isolation & purification; D011817:Rabbits; D011897:Random Allocation; D011994:Recombinant Proteins / Q000276:immunology; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000517:prevention & control*; D014614:Vaccines, Synthetic
D000909:Antibodies, Helminth; D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D010450:Endopeptidases; D000626:Aminopeptidases; C030596:hydrolase H