TY - JOUR
T1 - Low doses of flagellin-L2 multimer vaccines protect against challenge with diverse papillomavirus genotypes
AU - Kalnin, Kirill
AU - Tibbitts, Timothy
AU - Yan, Yanhua
AU - Stegalkina, Svetlana
AU - Shen, Lihua
AU - Costa, Victor
AU - Sabharwal, Robert
AU - Anderson, Stephen F.
AU - Day, Patricia M.
AU - Christensen, Neil
AU - Schiller, John T.
AU - Jagu, Subhashini
AU - Roden, Richard B.S.
AU - Almond, Jeffrey
AU - Kleanthous, Harold
N1 - Funding Information:
Conflict of interests : SJ and RBSR are co-inventors on L2 patents licensed to Shantha Biotechnics Ltd., GlaxoSmithKline, PaxVax Inc. and Acambis Inc., and have received grant support from Sanofi Pasteur. The terms of these arrangements are managed by Johns Hopkins University in accordance with its conflict of interest policies. JTS is a co-inventor on U.S. Government owed patents on papillomavirus L1 vaccines that are licensed to Merck & Co. and GlaxoSmithKline and L2 vaccines that are licensed to Shantha Biotechnics Ltd., GlaxoSmithKline, PaxVax Inc., Acambis Inc. and Sanofi Pasteur. Funding : This study was funded by Sanofi Pasteur . Ethical statement : Mouse studies were carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All animal studies were performed with the prior approval of the Animal Care and Use Committee of Johns Hopkins University (protocol MO08M19). Rabbit studies were performed according to the quality service and ethical treatment policy of Covance Inc. All Covance programs are AAALAC International accredited, and meet or exceed USDA Research License requirements, as well as those of the “Guide for Care and Use of Laboratory Animals”. All animal studies are performed with the prior approval of the Institutional Animal Care and Use Committee.
PY - 2014/6/12
Y1 - 2014/6/12
N2 - Genetically modified bacterial flagellin (Fla), a Toll-like receptor-5 (TLR5) ligand, was evaluated as a fusion partner for human papillomavirus (HPV) L2-based immunogens in two animal challenge models; either cutaneous inoculation of rabbits with HPV 'quasivirions' containing cottontail rabbit papillomavirus (CRPV) genomes that induce warts, or intra-vaginal inoculation of mice with HPV 'pseudovirions' encapsidating a luciferase reporter plasmid and measurement of bioluminescence to determine infectivity. An Escherichia coli production system was developed for flagellin-L2 (Fla-L2) fusions containing either monomeric HPV-16 L2 a.a. 11(× 11-200) or oligomeric L2 comprising a fusion of the a.a. 11-88 peptides of five (Fla~5 × 11-88) or eight (Fla~8 × 11-88) genital HPV types. Immunogenicity and bioactivity of Fla-L2 constructs were assessed using an in vitro neutralization and cell-based TLR-5 binding assay, respectively. Efficacy was evaluated following active immunization of rabbits or mice administered 3 intramuscular doses of Fla-L2 recombinants without exogenous adjuvant, followed by challenge. In addition, passive immunization studies of naïve rabbits with serial dilutions of pooled immune sera were used to determine End-Point Protection Titers (EPPT) for each formulation against a broader spectrum of HPV quasivirions. Efficacy was assessed for up to 10 weeks on the basis of wart volume induced following challenge and results compared to licensed L1-VLP vaccines (Gardasil and Cervarix). Following active immunization at doses as low as 1. μg, Fla-L2 fusions afforded complete protection against infection (mice) and disease (rabbits) following either homologous or heterologous HPV challenge. Passive immunization with anti-L2 immune sera discriminated between the different vaccine candidates under evaluation, demonstrated the protective role of antibody and suggested the superiority of this oligomeric L2-TLR5 agonist fusion approach compared to L1-based vaccines in its ability to cross-protect against non-vaccine HPV types.
AB - Genetically modified bacterial flagellin (Fla), a Toll-like receptor-5 (TLR5) ligand, was evaluated as a fusion partner for human papillomavirus (HPV) L2-based immunogens in two animal challenge models; either cutaneous inoculation of rabbits with HPV 'quasivirions' containing cottontail rabbit papillomavirus (CRPV) genomes that induce warts, or intra-vaginal inoculation of mice with HPV 'pseudovirions' encapsidating a luciferase reporter plasmid and measurement of bioluminescence to determine infectivity. An Escherichia coli production system was developed for flagellin-L2 (Fla-L2) fusions containing either monomeric HPV-16 L2 a.a. 11(× 11-200) or oligomeric L2 comprising a fusion of the a.a. 11-88 peptides of five (Fla~5 × 11-88) or eight (Fla~8 × 11-88) genital HPV types. Immunogenicity and bioactivity of Fla-L2 constructs were assessed using an in vitro neutralization and cell-based TLR-5 binding assay, respectively. Efficacy was evaluated following active immunization of rabbits or mice administered 3 intramuscular doses of Fla-L2 recombinants without exogenous adjuvant, followed by challenge. In addition, passive immunization studies of naïve rabbits with serial dilutions of pooled immune sera were used to determine End-Point Protection Titers (EPPT) for each formulation against a broader spectrum of HPV quasivirions. Efficacy was assessed for up to 10 weeks on the basis of wart volume induced following challenge and results compared to licensed L1-VLP vaccines (Gardasil and Cervarix). Following active immunization at doses as low as 1. μg, Fla-L2 fusions afforded complete protection against infection (mice) and disease (rabbits) following either homologous or heterologous HPV challenge. Passive immunization with anti-L2 immune sera discriminated between the different vaccine candidates under evaluation, demonstrated the protective role of antibody and suggested the superiority of this oligomeric L2-TLR5 agonist fusion approach compared to L1-based vaccines in its ability to cross-protect against non-vaccine HPV types.
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UR - http://www.scopus.com/inward/citedby.url?scp=84901475038&partnerID=8YFLogxK
U2 - 10.1016/j.vaccine.2014.04.032
DO - 10.1016/j.vaccine.2014.04.032
M3 - Article
C2 - 24780250
AN - SCOPUS:84901475038
SN - 0264-410X
VL - 32
SP - 3540
EP - 3547
JO - Vaccine
JF - Vaccine
IS - 28
ER -
