Generation of virus-like particles consisting of the major capsid protein VP1 of goose hemorrhagic polyomavirus and their application in serological tests.
Goose hemorrhagic polyomavirus (GHPV) is the causative agent of hemorrhagic nephritis and enteritis of geese (HNEG), a fatal disease of young geese with high mortality rates. GHPV cannot be efficiently propagated in tissue culture. To provide antigens for diagnostic tests and vaccines, its major structural protein VP1 was recombinantly expressed in Sf9 insect cells and in the yeast Saccharomyces cerevisiae. As demonstrated by density gradient centrifugation and electron microscopy, GHPV-VP1 expressed in insect cells formed virus-like particles (VLPs) with a diameter of 45 nm indistinguishable from infectious polyomavirus particles. However, efficiency of VLP formation was low as compared to the monkey polyomavirus SV-40-VP1. In yeast cells, GHPV-VP1 alone formed smaller VLPs, 20 nm in diameter. Remarkably, co-expression of GHPV-VP2 resulted in VLPs with a diameter of 45 nm. All three types of GHPV-VLPs were shown to hemagglutinate chicken erythrocytes. ELISA and hemagglutination inhibition tests using the VLPs as antigen detected GHPV-specific antibodies in up to 85.7% of sera derived from flocks with HNEG but in none of the sera of a clinically healthy flock. However, GHPV-specific antibodies were also detected in sera from two other flocks without HNEG indicating a broad distribution of GHPV due to subclinical or unrecognised infections.
Zielonka
Anja
A
Institute for Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany.
Gedvilaite
Alma
A
Ulrich
Rainer
R
Lüschow
Dörte
D
Sasnauskas
Kestutis
K
Müller
Hermann
H
Johne
Reimar
R
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000096:biosynthesis / Q000276:immunology; D001715:Bird Diseases / Q000097:blood* / Q000175:diagnosis; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D002460:Cell Line; D004751:Enteritis / Q000097:blood / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005777:Geese; D006385:Hemagglutination Inhibition Tests / Q000379:methods*; D007106:Immune Sera / Q000276:immunology; D007313:Insecta; D009393:Nephritis / Q000097:blood / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D011120:Polyomavirus / Q000737:chemistry* / Q000276:immunology; D027601:Polyomavirus Infections / Q000097:blood / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014412:Tumor Virus Infections / Q000097:blood / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D007106:Immune Sera; D011994:Recombinant Proteins; C062481:VP1 protein, polyomavirus