title: Preparation and properties of a papillomavirus infectious intermediate and its utility for neutralization studies.
abstract: We show that minor capsid protein L2 is full length in clinical virion isolates and prepare furin-cleaved pseudovirus (fcPsV) as a model of the infectious intermediate for multiple human papillomavirus (HPV) types. These fcPsV do not require furin for in vitro infection, and are fully infectious in vivo. Both the γ-secretase inhibitor XXI and carrageenan block fcPsV infection in vitro and in vivo implying that they act after furin-cleavage of L2. Despite their enhanced exposure of L2 epitopes, vaccination with fcPsV particles fails to induce L2 antibody, although L1-specific responses are similar to PsV with intact L2. FcPsV can be applied in a simple, high-throughput neutralization assay that detects L2-specific neutralizing antibodies with >10-fold enhanced sensitivity compared with the PsV-based assay. The PsV and fcPsV-based assays exhibit similar sensitivity for type-specific antibodies elicited by L1 virus-like particles (VLP), but the latter improves detection of L1-specific cross-type neutralizing antibodies.
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, MD 21231, USA.
authors.forename: Joshua W
authors.initials: JW
authors.lastname: Wang
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, MD 21231, USA.
authors.forename: Subhashini
authors.initials: S
authors.lastname: Jagu
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, MD 21231, USA.
authors.forename: Kihyuck
authors.initials: K
authors.lastname: Kwak
authors.affiliation: Department of Biostatistics, The Johns Hopkins University, Baltimore, MD 21231, USA.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Oncology, The Johns Hopkins University, Baltimore, MD 21231, USA.
authors.forename: Shiwen
authors.initials: S
authors.lastname: Peng
authors.affiliation: Laboratory of Viral Oncology, Division of Immunology, Allergy and Infectious Diseases, Department of Dermatology, Medical University Vienna (MUW), Vienna, Austria.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kirnbauer
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, MD 21231, USA; Department of Oncology, The Johns Hopkins University, Baltimore, MD 21231, USA; Department of Gynecology and Obstetrics, The Johns Hopkins University, Baltimore, MD 21231, USA. Electronic address: roden@jhmi.edu.
authors.forename: Richard B S
authors.initials: RB
authors.lastname: Roden
mesh_terms: D052685:Alphapapillomavirus / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D036022:Capsid Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D003429:Cross Reactions; D045683:Furin / Q000378:metabolism; D006801:Humans; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D030361:Papillomavirus Infections / Q000276:immunology / Q000517:prevention & control / Q000821:virology*; D053918:Papillomavirus Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D036022:Capsid Proteins; D053918:Papillomavirus Vaccines; D045683:Furin
keywords: Carrageenan; Furin; Gamma secretase; Heparin; Human papillomavirus (HPV); Infectious intermediate; L2; Minor capsid protein; Neutralization; Papillomavirus