Chimeric L2-Based Virus-Like Particle (VLP) Vaccines Targeting Cutaneous Human Papillomaviruses (HPV)

Common cutaneous human papillomavirus (HPV) types induce skin warts, whereas species beta HPV are implicated, together with UV-radiation, in the development of non-melanoma skin cancer (NMSC) in immunosuppressed patients. Licensed HPV vaccines contain virus-like particles (VLP) self-assembled from L1 major capsid proteins that provide type-restricted protection against mucosal HPV infections causing cervical and other ano-genital and oro-pharyngeal carcinomas and warts (condylomas), but do not target heterologous HPV. Experimental papillomavirus vaccines have been designed based on L2 minor capsid proteins that contain type-common neutralization epitopes, to broaden protection to heterologous mucosal and cutaneous HPV types. Repetitive display of the HPV16 L2 cross-neutralization epitope RG1 (amino acids (aa) 17–36) on the surface of HPV16 L1 VLP has greatly enhanced immunogenicity of the L2 peptide. To more directly target cutaneous HPV, L1 fusion proteins were designed that incorporate the RG1 homolog of beta HPV17, the beta HPV5 L2 peptide aa53-72, or the common cutaneous HPV4 RG1 homolog, inserted into DE surface loops of HPV1, 5, 16 or 18 L1 VLP scaffolds. Baculovirus expressed chimeric proteins self-assembled into VLP and VLP-raised NZW rabbit immune sera were evaluated by ELISA and L1- and L2-based pseudovirion (PsV) neutralizing assays, including 12 novel beta PsV types. Chimeric VLP displaying the HPV17 RG1 epitope, but not the HPV5L2 aa53-72 epitope, induced cross-neutralizing humoral immune responses to beta HPV. In vivo cross-protection was evaluated by passive serum transfer in a murine PsV challenge model. Immune sera to HPV16L1-17RG1 VLP (cross-) protected against beta HPV5/20/24/38/96/16 (but not type 76), while antisera to HPV5L1-17RG1 VLP cross-protected against HPV20/24/96 only, and sera to HPV1L1-4RG1 VLP cross-protected against HPV4 challenge. In conclusion, RG1-based VLP are promising next generation vaccine candidates to target cutaneous HPV infections.

[1]  R. Roden,et al.  Developments in L2-based human papillomavirus (HPV) vaccines. , 2017, Virus research.

[2]  R. Shoemaker,et al.  426 cGMP production of a chimeric virus-like particle vaccine (RG1-VLP) for prevention of HPV-associated cancers , 2016 .

[3]  N. Trinklein,et al.  A Single Human Papillomavirus Vaccine Dose Improves B Cell Memory in Previously Infected Subjects , 2016, EBioMedicine.

[4]  M. Boxus,et al.  Broad Cross-Protection Is Induced in Preclinical Models by a Human Papillomavirus Vaccine Composed of L1/L2 Chimeric Virus-Like Particles , 2016, Journal of Virology.

[5]  J. Tomimori,et al.  Human papillomavirus detected in viral warts of renal transplant recipients , 2016, Transplant infectious disease : an official journal of the Transplantation Society.

[6]  R. Roden,et al.  Capsid display of a conserved human papillomavirus L2 peptide in the adenovirus 5 hexon protein: a candidate prophylactic hpv vaccine approach , 2015, Virology Journal.

[7]  K. Egawa,et al.  Human Papillomaviruses; Epithelial Tropisms, and the Development of Neoplasia , 2015, Viruses.

[8]  D. Peabody,et al.  Preclinical refinements of a broadly protective VLP-based HPV vaccine targeting the minor capsid protein, L2. , 2015, Vaccine.

[9]  S. Shafti-Keramat,et al.  A Chimeric 18L1-45RG1 Virus-Like Particle Vaccine Cross-Protects against Oncogenic Alpha-7 Human Papillomavirus Types , 2015, PloS one.

[10]  S. Tyring,et al.  HPV Carcinomas in Immunocompromised Patients , 2015, Journal of clinical medicine.

[11]  M. Feltkamp,et al.  Human Beta‐papillomavirus infection and keratinocyte carcinomas , 2015, The Journal of pathology.

[12]  M. Poljak,et al.  Betapapillomaviruses in the anal canal of HIV positive and HIV negative men who have sex with men , 2014, BMC Infectious Diseases.

[13]  R. Roden,et al.  Low doses of flagellin-L2 multimer vaccines protect against challenge with diverse papillomavirus genotypes. , 2014, Vaccine.

[14]  R. Roden,et al.  Impact of Inhibitors and L2 Antibodies upon the Infectivity of Diverse Alpha and Beta Human Papillomavirus Types , 2014, PloS one.

[15]  M. Storgaard,et al.  Comparison of the immunogenicity of Cervarix® and Gardasil® human papillomavirus vaccines for oncogenic non-vaccine serotypes HPV-31, HPV-33, and HPV-45 in HIV-infected adults , 2014, Human vaccines & immunotherapeutics.

[16]  F. Rösl,et al.  Protective Vaccination against Papillomavirus-Induced Skin Tumors under Immunocompetent and Immunosuppressive Conditions: A Preclinical Study Using a Natural Outbred Animal Model , 2014, PLoS pathogens.

[17]  Neil D. Christensen,et al.  A Cryo-Electron Microscopy Study Identifies the Complete H16.V5 Epitope and Reveals Global Conformational Changes Initiated by Binding of the Neutralizing Antibody Fragment , 2014, Journal of Virology.

[18]  L. Denny,et al.  Safety and immunogenicity of the HPV-16/18 AS04-adjuvanted vaccine in HIV-positive women in South Africa: a partially-blind randomised placebo-controlled study. , 2013, Vaccine.

[19]  D. Peabody,et al.  A universal virus-like particle-based vaccine for human papillomavirus: longevity of protection and role of endogenous and exogenous adjuvants. , 2013, Vaccine.

[20]  Deepali Kumar,et al.  Immunogenicity of Quadrivalent Human Papillomavirus Vaccine in Organ Transplant Recipients , 2013, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[21]  J. Dillner,et al.  Efficacy of RG1-VLP Vaccination against Infections with Genital and Cutaneous Human Papillomaviruses , 2013, The Journal of investigative dermatology.

[22]  R. Roden,et al.  Optimization of Multimeric Human Papillomavirus L2 Vaccines , 2013, PloS one.

[23]  Steven M Holland,et al.  Warts and all: human papillomavirus in primary immunodeficiencies. , 2012, The Journal of allergy and clinical immunology.

[24]  D. Lowy,et al.  A Human Papillomavirus (HPV) In Vitro Neutralization Assay That Recapitulates the In Vitro Process of Infection Provides a Sensitive Measure of HPV L2 Infection-Inhibiting Antibodies , 2012, Clinical and Vaccine Immunology.

[25]  Jong-Sup Park,et al.  Oral administration of HPV-16 L2 displayed on Lactobacillus casei induces systematic and mucosal cross-neutralizing effects in Balb/c mice. , 2012, Vaccine.

[26]  J. Kleinschmidt,et al.  Intranasal vaccination with AAV5 and 9 vectors against human papillomavirus type 16 in rhesus macaques. , 2012, Human gene therapy.

[27]  J. Bordeaux,et al.  The high risk HPV16 L2 minor capsid protein has multiple transport signals that mediate its nucleocytoplasmic traffic. , 2012, Virology.

[28]  J. Cardoso,et al.  Cutaneous manifestations of human papillomaviruses: a review. , 2011, Acta dermatovenerologica Alpina, Pannonica, et Adriatica.

[29]  R. Burk,et al.  The oral cavity contains abundant known and novel human papillomaviruses from the Betapapillomavirus and Gammapapillomavirus genera. , 2011, The Journal of infectious diseases.

[30]  D. Peabody,et al.  A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2 , 2011, PloS one.

[31]  A. Moscicki,et al.  Safety and Immunogenicity of a Quadrivalent Human Papillomavirus (Types 6, 11, 16, and 18) Vaccine in HIV-Infected Children 7 to 12 Years Old , 2010, Journal of acquired immune deficiency syndromes.

[32]  R. Roden,et al.  Vaccination with multimeric L2 fusion protein and L1 VLP or capsomeres to broaden protection against HPV infection. , 2010, Vaccine.

[33]  E. de Villiers,et al.  Classification of papillomaviruses (PVs) based on 189 PV types and proposal of taxonomic amendments. , 2010, Virology.

[34]  Abdelatif Elouahabi,et al.  Molecular and structural characterization of the L1 virus-like particles that are used as vaccine antigens in Cervarix™, the AS04-adjuvanted HPV-16 and -18 cervical cancer vaccine , 2010, Human vaccines.

[35]  Sonja Textor,et al.  Virus-like particles and capsomeres are potent vaccines against cutaneous alpha HPVs. , 2010, Vaccine.

[36]  R. Roden,et al.  Chimeric L1-L2 Virus-Like Particles as Potential Broad-Spectrum Human Papillomavirus Vaccines , 2009, Journal of Virology.

[37]  C. Kovarik,et al.  Dermatologic manifestations of HPV in HIV-infected individuals , 2009, Current HIV/AIDS reports.

[38]  R. Herd,et al.  Presence of beta human papillomaviruses in nonmelanoma skin cancer from organ transplant recipients and immunocompetent patients in the West of Scotland , 2009, The British journal of dermatology.

[39]  D. Lowy,et al.  Concatenated Multitype L2 Fusion Proteins as Candidate Prophylactic Pan-Human Papillomavirus Vaccines , 2009, Journal of the National Cancer Institute.

[40]  Y. Jacob,et al.  The EVER Proteins as a Natural Barrier against Papillomaviruses: a New Insight into the Pathogenesis of Human Papillomavirus Infections , 2009, Microbiology and Molecular Biology Reviews.

[41]  S. Ottonello,et al.  Potent anti-HPV immune responses induced by tandem repeats of the HPV16 L2 (20 -- 38) peptide displayed on bacterial thioredoxin. , 2009, Vaccine.

[42]  D. Lowy,et al.  Concatenated Multitype L2 Fusion Proteins as Candidate Prophylactic Pan-Human Papillomavirus Vaccines , 2009, Journal of the National Cancer Institute.

[43]  M. Favre,et al.  Serological relationship between cutaneous human papillomavirus types 5, 8 and 92. , 2009, The Journal of general virology.

[44]  R. Roden,et al.  Protection against heterologous human papillomavirus challenge by a synthetic lipopeptide vaccine containing a broadly cross-neutralizing epitope of L2 , 2008, Proceedings of the National Academy of Sciences.

[45]  M. Favre,et al.  Serological relationship between cutaneous human papillomavirus types 5, 8 and 92 , 2008 .

[46]  C. Buck,et al.  Production of Papillomavirus‐Based Gene Transfer Vectors , 2007, Current protocols in cell biology.

[47]  I. Bossis,et al.  A Protective and Broadly Cross-Neutralizing Epitope of Human Papillomavirus L2 , 2007, Journal of Virology.

[48]  R. Roden,et al.  Protection of Rabbits against Challenge with Rabbit Papillomaviruses by Immunization with the N Terminus of Human Papillomavirus Type 16 Minor Capsid Antigen L2 , 2007, Journal of Virology.

[49]  P. Choyke,et al.  Genital transmission of HPV in a mouse model is potentiated by nonoxynol-9 and inhibited by carrageenan , 2007, Nature Medicine.

[50]  Y. Ishii,et al.  Neutralization of HPV16, 18, 31, and 58 pseudovirions with antisera induced by immunizing rabbits with synthetic peptides representing segments of the HPV16 minor capsid protein L2 surface region. , 2007, Virology.

[51]  R. Roden,et al.  How will HPV vaccines affect cervical cancer? , 2006, Nature Reviews Cancer.

[52]  Luise Florin,et al.  Identification of a Dynein Interacting Domain in the Papillomavirus Minor Capsid Protein L2 , 2006, Journal of Virology.

[53]  A. Green,et al.  Markers of Cutaneous Human Papillomavirus Infection in Individuals with Tumor-Free Skin, Actinic Keratoses, and Squamous Cell Carcinoma , 2006, Cancer Epidemiology Biomarkers & Prevention.

[54]  K. Palmer,et al.  Modified tobacco mosaic virus particles as scaffolds for display of protein antigens for vaccine applications. , 2006, Virology.

[55]  D. Lowy,et al.  Cross-neutralization of cutaneous and mucosal Papillomavirus types with anti-sera to the amino terminus of L2. , 2005, Virology.

[56]  C. Proby,et al.  Human papillomavirus-DNA loads in actinic keratoses exceed those in non-melanoma skin cancers. , 2005, The Journal of investigative dermatology.

[57]  J. Moroianu,et al.  The L2 Minor Capsid Protein of Human Papillomavirus Type 16 Interacts with a Network of Nuclear Import Receptors , 2004, Journal of Virology.

[58]  D. Lowy,et al.  Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. , 2004, Virology.

[59]  D. Lowy,et al.  Efficient Intracellular Assembly of Papillomaviral Vectors , 2004, Journal of Virology.

[60]  D. Galloway,et al.  Identification of a Human Papillomavirus Type 16-Specific Epitope on the C-Terminal Arm of the Major Capsid Protein L 1 , 2003 .

[61]  H. Pfister.  Chapter 8: Human papillomavirus and skin cancer. , 2003, Journal of the National Cancer Institute. Monographs.

[62]  N. Ramoz,et al.  Mutations in two adjacent novel genes are associated with epidermodysplasia verruciformis , 2002, Nature Genetics.

[63]  W. Mcclements,et al.  Hybrid papillomavirus L1 molecules assemble into virus-like particles that reconstitute conformational epitopes and induce neutralizing antibodies to distinct HPV types. , 2001, Virology.

[64]  J. Bouwes Bavinck,et al.  Human papillomavirus infection and skin cancer risk in organ transplant recipients. , 2001, The journal of investigative dermatology. Symposium proceedings.

[65]  Y. Taketani,et al.  Human Papillomavirus Type 16 Minor Capsid Protein L2 N-Terminal Region Containing a Common Neutralization Epitope Binds to the Cell Surface and Enters the Cytoplasm , 2001, Journal of Virology.

[66]  J. McGregor,et al.  Human papillomavirus infection and non‐melanoma skin cancer in immunosuppressed and immunocompetent individuals , 2000, Journal of medical virology.

[67]  Y. Taketani,et al.  Common Neutralization Epitope in Minor Capsid Protein L2 of Human Papillomavirus Types 16 and 6 , 1999, Journal of Virology.

[68]  Y. Taketani,et al.  A surface immunodeterminant of human papillomavirus type 16 minor capsid protein L2. , 1998, Virology.

[69]  P. Barbosa,et al.  Epidemiology of plantar verrucae in HIV-infected individuals. , 1998, Journal of Acquired Immune Deficiency Syndromes & Human Retrovirology.

[70]  J. Dillner,et al.  A monoclonal antibody against intact human papillomavirus type 16 capsids blocks the serological reactivity of most human sera. , 1997, The Journal of general virology.

[71]  D. Lowy,et al.  Characterization of a human papillomavirus type 16 variant-dependent neutralizing epitope , 1997, Journal of virology.

[72]  J. Palefsky.  Cutaneous and genital HPV-associated lesions in HIV-infected patients. , 1997, Clinics in dermatology.

[73]  J. Dillner,et al.  Surface conformational and linear epitopes on HPV-16 and HPV-18 L1 virus-like particles as defined by monoclonal antibodies. , 1996, Virology.

[74]  G. Grindlay,et al.  Vaccination of cattle with bovine papillomavirus type 4 L2 elicits the production of virus-neutralizing antibodies. , 1996, The Journal of general virology.

[75]  D. Lowy,et al.  Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles , 1993, Journal of virology.

[76]  D. Lowy,et al.  Papillomavirus L1 major capsid protein self-assembles into virus-like particles that are highly immunogenic. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[77]  M. Stanley,et al.  Production and characterisation of a monoclonal antibody to human papillomavirus type 16 using recombinant vaccinia virus. , 1990, Journal of clinical pathology.

[78]  W. Lancaster,et al.  Monoclonal antibodies to genus- and type-specific papillomavirus structural antigens. , 1986, Intervirology.

[79]  S. Jablonska,et al.  Epidermodysplasia verruciformis as a model in studies on the role of papovaviruses in oncogenesis. , 1972, Cancer research.