Expression of human papillomavirus type 11 L1 protein in insect cells: in vivo and in vitro assembly of viruslike particles.
The L1 coat protein of human papillomavirus type 11 (HPV-11) was expressed in Sf-9 insect cells with the recombinant baculovirus vector Ac11L1. Viruslike particles (VLPs) were identified by electron microscopy in the nucleus and cytoplasm of Sf-9 cells infected with Ac11L1. The L1 protein was purified from Ac11L1-infected insect cells. The purified protein spontaneously assembled in vitro into various aggregates, including particles appearing similar to empty virions. Reaction of VLP-containing insect cell extracts with antisera directed against either denatured or nondenatured capsid epitopes in Western blot (immunoblot) and immuno-dot blot assays suggested that conformational epitopes present in native HPV-11 infectious virions were also present on the baculovirus-produced HPV-11 VLPs. Immuno-dot blot assays using human sera obtained from individuals with biopsy-proven condyloma acuminatum correlated closely with results previously obtained in HPV-11 whole virus particle-based enzyme-linked immunosorbent assays. These morphologic and immunologic similarities to native HPV-11 virions suggest that recombinant VLPs produced in the baculovirus system may be useful in seroepidemiology and pathogenesis studies of genital HPV infection and that they may also be potential candidates for vaccine development.
Department of Medicine, University of Rochester School of Medicine and Dentistry, New York 14642.
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Rose
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Bonnez
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Reichman
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Garcea
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D016367:Baculoviridae; D001483:Base Sequence; D002213:Capsid / Q000235:genetics* / Q000276:immunology; D036022:Capsid Proteins; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D066298:In Vitro Techniques; D007915:Lepidoptera; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000276:immunology; D027383:Papillomaviridae / Q000235:genetics* / Q000254:growth & development / Q000276:immunology; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D014771:Virion / Q000648:ultrastructure; D014779:Virus Replication
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C080100:oncogene viral capsid protein L2, human papillomavirus type 33