title: Hybrid papillomavirus L1 molecules assemble into virus-like particles that reconstitute conformational epitopes and induce neutralizing antibodies to distinct HPV types.
abstract: Human papillomavirus (HPV) hybrid virus-like particles (VLPs) were prepared using complementary regions of the major capsid L1 proteins of HPV-11 and -16. These hybrid L1 proteins were tested for assembly into VLPs, for presentation and mapping of conformational neutralizing epitopes, and as immunogens in rabbits and mice. Two small noncontiguous hypervariable regions of HPV-16 L1, when replaced into the HPV-11 L1 backbone, produced an assembly-positive hybrid L1 which was recognized by the type-specific, conformationally dependent HPV-16 neutralizing monoclonal antibody (N-MAb) H16.V5. Several new N-MAbs that were generated following immunization of mice with wild-type HPV-16 L1 VLPs also recognized this reconstructed VLP, demonstrating that these two hypervariable regions collectively constituted an immunodominant epitope. When a set of hybrid VLPs was tested as immunogens in rabbits, antibodies to both HPV-11 and -16 wild-type L1 VLPs were obtained. One of the hybrid VLPs containing hypervariable FG and HI loops of HPV-16 L1 replaced into an HPV-11 L1 background provoked neutralizing activity against both HPV-11 and HPV-16. In addition, conformationally dependent and type-specific MAbs to both HPV-11 and HPV-16 L1 VLP were obtained from mice immunized with hybrid L1 VLPs. These data indicated that hybrid L1 proteins can be constructed that retain VLP-assembly properties, retain type-specific conformational neutralizing epitopes, can map noncontiguous regions of L1 which constitute type-specific conformational neutralizing epitopes recognized by N-MAbs, and trigger polyclonal antibodies which can neutralize antigenically unrelated HPV types.
authors.affiliation: The Jake Gittlen Cancer Research Institute, Pathology Department, Milton S. Hershey Medical Center, 500 University Drive, Hershey, PA 17033, USA. ndc1@psu.edu
authors.forename: N D
authors.initials: ND
authors.lastname: Christensen
authors.forename: N M
authors.initials: NM
authors.lastname: Cladel
authors.forename: C A
authors.initials: CA
authors.lastname: Reed
authors.forename: L R
authors.initials: LR
authors.lastname: Budgeon
authors.forename: M E
authors.initials: ME
authors.lastname: Embers
authors.forename: D M
authors.initials: DM
authors.lastname: Skulsky
authors.forename: W L
authors.initials: WL
authors.lastname: McClements
authors.forename: S W
authors.initials: SW
authors.lastname: Ludmerer
authors.forename: K U
authors.initials: KU
authors.lastname: Jansen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis; D036022:Capsid Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D051379:Mice; D009500:Neutralization Tests; D009856:Oncogene Proteins, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D027383:Papillomaviridae / Q000235:genetics / Q000276:immunology*; D011487:Protein Conformation; D011817:Rabbits; D014771:Virion
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; C094776:L1 protein, Human papillomavirus type 11; D009856:Oncogene Proteins, Viral; C063317:L1 protein, Human papillomavirus type 16