title: Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles.
abstract: The L1 genes of two human papillomavirus type 16 (HPV16) isolates derived from condylomata acuminata were used to express the L1 major capsid protein in insect cells via recombinant baculoviruses. Both L1 major capsid proteins self-assembled into virus-like particles (VLP) with high efficiency and could be purified in preparative amounts on density gradients. The yield of VLP was 3 orders of magnitude higher than what has been obtained previously, using L1 derived from the prototype HPV16. DNA sequence comparison identified a single nonconserved amino acid change to be responsible for the inefficient self-assembly of the prototype L1. VLP were also obtained by expressing L1 of HPV6, HPV11, and cottontail rabbit papillomavirus, indicating that L1 from a variety of papillomaviruses has the intrinsic capacity to self-assemble into VLP. Coexpression of HPV16 L1 plus L2 by using a baculovirus double-expression vector also resulted in efficient self-assembly of VLP, and the average particle yield increased about fourfold in comparison to when L1 only was expressed. Coimmunoprecipitation of L1 and L2 and cosedimentation of the two proteins in a sucrose gradient demonstrated that L2 was incorporated into the particles. The ability to generate preparative amounts of HPV16 L1 and L1-L2 VLP may have implications for the development of a serological assay to detect anti-HPV16 virion immune responses to conformational epitopes and for immunoprophylaxis against HPV16 infection.
authors.affiliation: Laboratory of Cellular Oncology, National Cancer Institute, Bethesda, Maryland 20892.
authors.forename: R
authors.initials: R
authors.lastname: Kirnbauer
authors.forename: J
authors.initials: J
authors.lastname: Taub
authors.forename: H
authors.initials: H
authors.lastname: Greenstone
authors.forename: R
authors.initials: R
authors.lastname: Roden
authors.forename: M
authors.initials: M
authors.lastname: Dürst
authors.forename: L
authors.initials: L
authors.lastname: Gissmann
authors.forename: D R
authors.initials: DR
authors.lastname: Lowy
authors.forename: J T
authors.initials: JT
authors.lastname: Schiller
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002478:Cells, Cultured; D003218:Condylomata Acuminata / Q000382:microbiology; D005814:Genes, Viral; D006801:Humans; D008969:Molecular Sequence Data; D009024:Morphogenesis; D009036:Moths / Q000166:cytology; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism*; D027383:Papillomaviridae / Q000254:growth & development* / Q000648:ultrastructure; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000378:metabolism; D017421:Sequence Analysis
chemical_list: D036022:Capsid Proteins; C063316:L2 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16