Hybrid N-glycans on the host protective activation-associated secreted proteins of Ostertagia ostertagi and their importance in immunogenicity.
Previous vaccination trials with calves have shown that intramuscular immunization with natively purified activation-associated secreted proteins (ASPs) of Ostertagia ostertagi induces protection against a homologous challenge infection with a 74% reduction in cumulative faecal egg counts and a significant reduction in worm length. Recently, O. ostertagi ASP1 was recombinantly expressed using a baculovirus system and tested in a vaccination trial. However, immunized calves failed to recognize native ASPs and no protection was observed. These results suggest an important structural difference between the baculo r-ASP1 and its native counterpart. Therefore, we investigated whether glycans and/or structural epitopes are key features in the induction of a protective immune response. The results show that ASPs carry two hybrid N-glycosylations with a complex alpha-1,3-arm, an unprocessed alpha-1,6-arm and an alpha-1,6-fucose core. While removal of these glycans had little effect on antibody recognition by vaccinated animals, denaturing and reducing the proteins dramatically reduced recognition, suggesting the importance of conformational protein backbone epitopes.
Meyvis
Yves
Y
Laboratory of Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium. yves.meyvis@ugent.be
Callewaert
Nico
N
Gevaert
Kris
K
Timmerman
Evy
E
Van Durme
Joost
J
Schymkowitz
Joost
J
Rousseau
Frederic
F
Vercruysse
Jozef
J
Claerebout
Edwin
E
Geldhof
Peter
P
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000378:metabolism; D000947:Antigens, Helminth / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D010028:Ostertagia / Q000737:chemistry* / Q000276:immunology*; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000032:analysis* / Q000276:immunology*; D011485:Protein Binding; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D011134:Polysaccharides; D011994:Recombinant Proteins