title: Seroepidemiology of Human Papillomavirus 16 (HPV16) L2 and Generation of L2-Specific Human Chimeric Monoclonal Antibodies.
abstract: Presently, the seroprevalence of human papillomavirus (HPV) minor capsid antigen L2-reactive antibody is not well understood, and no serologic standard exists for L2-specific neutralizing antibodies. Therefore, we screened a total of 1,078 serum samples for HPV16 L2 reactivity, and these were obtained from four prior clinical studies: a population-based (n = 880) surveillance study with a high-risk HPV DNA prevalence of 10.8%, a cohort study of women (n = 160) with high-grade cervical intraepithelial neoplasia (CIN), and two phase II trials in women with high-grade vulvar intraepithelial neoplasia (VIN) receiving imiquimod therapy combined with either photodynamic therapy (PDT) (n = 19) or vaccination with a fusion protein comprising HPV16 L2, E7, and E6 (TA-CIN) (n = 19). Sera were screened sequentially by HPV16 L2 enzyme-linked immunosorbent assay (ELISA) and then Western blot. Seven of the 1,078 serum samples tested had L2-specific antibodies, but none were detectably neutralizing for HPV16. To develop a standard, we substituted human IgG1 sequences into conserved regions of two rodent monoclonal antibodies (MAbs) specific for neutralizing epitopes at HPV16 L2 residues 17 to 36 and 58 to 64, creating JWW-1 and JWW-2, respectively. These chimeric MAbs retained neutralizing activity and together reacted with 33/34 clinically relevant HPV types tested. In conclusion, our inability to identify an HPV16 L2-specific neutralizing antibody response even in the sera of patients with active genital HPV disease suggests the subdominance of L2 protective epitopes and the value of the chimeric MAbs JWW-1 and JWW-2 as standards for immunoassays to measure L2-specific human antibodies.
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Joshua W
authors.initials: JW
authors.lastname: Wang
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Subhashini
authors.initials: S
authors.lastname: Jagu
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Wai-Hong
authors.initials: WH
authors.lastname: Wu
authors.affiliation: Department of Pediatrics, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Raphael P
authors.initials: RP
authors.lastname: Viscidi
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Anne
authors.initials: A
authors.lastname: Macgregor-Das
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Jessica M
authors.initials: JM
authors.lastname: Fogel
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Kihyuck
authors.initials: K
authors.lastname: Kwak
authors.affiliation: Woman's Cancer Centre, St. Mary's Hospital, University of Manchester, Manchester, United Kingdom.
authors.forename: Sai
authors.initials: S
authors.lastname: Daayana
authors.affiliation: Woman's Cancer Centre, St. Mary's Hospital, University of Manchester, Manchester, United Kingdom.
authors.forename: Henry
authors.initials: H
authors.lastname: Kitchener
authors.affiliation: Paterson Building Institute of Cancer Sciences, University of Manchester, Manchester, United Kingdom.
authors.forename: Peter L
authors.initials: PL
authors.lastname: Stern
authors.affiliation: Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico, USA.
authors.forename: Patti E
authors.initials: PE
authors.lastname: Gravitt
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA Department of Oncology, The Johns Hopkins University, Baltimore, Maryland, USA Department of Gynecology and Obstetrics, The Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Cornelia L
authors.initials: CL
authors.lastname: Trimble
authors.affiliation: Department of Pathology, The Johns Hopkins University, Baltimore, Maryland, USA Department of Oncology, The Johns Hopkins University, Baltimore, Maryland, USA Department of Gynecology and Obstetrics, The Johns Hopkins University, Baltimore, Maryland, USA roden@jhmi.edu.
authors.forename: Richard B S
authors.identifier: http://orcid.org/0000-0002-2506-0195
authors.initials: RB
authors.lastname: Roden
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology* / Q000302:isolation & purification*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology*; D015331:Cohort Studies; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D052162:Human papillomavirus 16 / Q000276:immunology*; D006801:Humans; D008875:Middle Aged; D009500:Neutralization Tests; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D030361:Papillomavirus Infections / Q000453:epidemiology / Q000276:immunology / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D016036:Seroepidemiologic Studies; D012698:Serologic Tests / Q000379:methods / Q000592:standards
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; C063316:L2 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins