Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Oct:575:63-73.
doi: 10.1016/j.virol.2022.08.006. Epub 2022 Aug 23.

Vaccination with human alphapapillomavirus-derived L2 multimer protects against human betapapillomavirus challenge, including in epidermodysplasia verruciformis model mice

Pola Olczak  1 , Margaret Wong  1 , Hua-Ling Tsai  2 , Hao Wang  2 , Reinhard Kirnbauer  3 , Andrew J Griffith  4 , Paul F Lambert  5 , Richard Roden  6
Affiliations

Vaccination with human alphapapillomavirus-derived L2 multimer protects against human betapapillomavirus challenge, including in epidermodysplasia verruciformis model mice

Pola Olczak et al. Virology. 2022 Oct.

Abstract

Human alphapapillomaviruses (αHPV) infect genital mucosa, and a high-risk subset is a necessary cause of cervical cancer. Licensed L1 virus-like particle (VLP) vaccines offer immunity against the nine most common αHPV associated with cervical cancer and genital warts. However, vaccination with an αHPV L2-based multimer vaccine, α11-88x5, protected mice and rabbits from vaginal and skin challenge with diverse αHPV types. While generally clinically inapparent, human betapapillomaviruses (βHPV) are possibly associated with cutaneous squamous cell carcinoma (CSCC) in epidermodysplasia verruciformis (EV) and immunocompromised patients. Here we show that α11-88x5 vaccination protected wild type and EV model mice against HPV5 challenge. Passive transfer of antiserum conferred protection independently of Fc receptors (FcR) or Gr-1+ phagocytes. Antisera demonstrated robust antibody titers against ten βHPV by L1/L2 VLP ELISA and neutralized and protected against challenge by 3 additional βHPV (HPV49/76/96). Thus, unlike the licensed vaccines, α11-88x5 vaccination elicits broad immunity against αHPV and βHPV.

Keywords: Cutaneous squamous cell carcinoma; Epidermodysplasia verruciformis; Fc receptor; HPV; Human papillomaviruses; L1 capsid protein; L2 capsid protein; Multimer; Preventive vaccination; Virus-like particle.

PubMed Disclaimer

Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Under license agreements between BravoVax Co., Ltd. and the Johns Hopkins University, Dr. Roden is entitled to distributions of payments associated with an invention described in this publication. Dr. Roden also owns equity in PathoVax LLC, Up Therapeutics LLC, Papivax LLC and Papivax Biotech Inc. and is a member of their scientific advisory boards. These arrangements have been reviewed and approved by the Johns Hopkins University in accordance with its conflict of interest policies.

Figures

Figure 1.
Figure 1.. Design of α11-88x5 fusion protein vaccine and antibody responses elicited to L1/L2 VLP.
(A) Sequence alignment of five HPV types included in the α11-88x5 vaccine. (B) Sequence alignment of the papillomavirus types the multimer vaccine was assessed against in the current study. Sequence alignments were generated using Jalview 2.11.1.7. (http://www.jalview.org/). Color scheme of amino acids corresponds to a default ClustalX setting. When the percentage threshold of amino acid per column was met, coloring reflects the following characteristics of amino acids: blue (hydrophobic), red (positive charge), magenta (negative charge), green (polar), pink (cysteines), orange (glycines), yellow (prolines), cyan (aromatic), white (unconserved). (C) Virus-like particles (VLP) used as antigens for the ELISA assays were produced in 293EXPI cells using mammalian expression system and purified by density gradient. Preparations (1 μg of L1) of each HPV VLP were analyzed by SDS-PAGE and SimplyBlue SafeStain (Thermo Fisher). (D) Expression of L2 therein was assessed by Western blot analysis using rabbit antiserum to α11-88x5. (E) Sera collected two weeks after 10 mice/group were vaccinated three times at 2 week intervals with 25 μg α11-88x5 on 50 μg aluminum phosphate (blue dots) or Gardasil®9 (1/20th human dose) (black dots), were analyzed by direct ELISA using α11-88x5 or the indicated papillomavirus L1 or L1/L2 VLP as antigens. Scatter plots indicate individual serum titers, while the bars indicate the mean values and error bars show standard deviation.
Figure 2
Figure 2. Vaccination with α11-88x5 protects against HPV5 PsV challenge in wild type and EV model mice.
Female FVB WT (n=10 or 8/group) or Tmc6Δ/Δ (n=10/group) or Tmc8Δ/Δ (n=5/group) mice were immunized with 25 μg of multimer vaccine formulated on 50 μg aluminum phosphate (alum) or alum adjuvant only, 3 times at two-week intervals. Two weeks post final vaccination, the mice were challenged with HPV5 PsV. P values were calculated by ordinary one-way ANOVA with the Šídák’s multiple comparisons test.
Figure 3.
Figure 3.. Assessment of protection against challenge with HPV49, 76 and 96 pseudovirions after passive and active vaccination.
(A) Protection via passive transfer of rabbit antisera to Gardasil®9 or α11-88x5 (50 μg dose on Freunds adjuvant) was assessed in CD-1 ISG mice (n=5/group) followed by vaginal challenge with βHPV PsVs derived from types 49, 76 and 96. Fifty microliters of sera from individual rabbits were passively transferred each mouse by i.p. injection. Mice were challenged intravaginally with PsV the following day. (B) Female CD-1 IGS mice (n=10/group) were immunized with multimer vaccine (25 μg formulated on alum), Gardasil ®9, or alum adjuvant by i.m. injection three times in 2 week intervals. Two weeks post final vaccination, the mice were challenged intravaginally with βHPV PsVs derived from types 49, 76 and 96. The bioluminescence images were acquired 72 h post challenge for 5 min with a Xenogen IVIS 100. P values were calculated by ordinary one-way ANOVA with the Dunnett’s multiple comparisons test.
Figure 4.
Figure 4.. Impact of dose, or FcγR knockout and Gr-1 antibody depletion in the recipient upon protection from HPV5 challenge via passive transfer of α11-88x5 antiserum.
Protection via passive transfer of pooled sera from mice vaccinated with 25 μg of α11-88x5 multimer vaccine formulated on 50 μg aluminum phosphate, or alum only, was assessed in mice following intravaginal challenge with HPV5 PsV. (A) Either 3 μL, 10 μL, 33 μL or 100 μL of pooled α11-88x5 antisera or 100 μL of Cervarix® or alum only pooled mouse antisera or 33 μL HPV5 L1 VLP antiserum from a rabbit were administered i.p. into individual female CD-1 ISG mice followed by intravaginal HPV5 PsV challenge the following day. The bioluminescence images were acquired 72 h post challenge for 5 min with a Xenogen IVIS 100 and quantified in the region of interest (photons/sec) or an irrelevant region of the same size to determine background. The lines indicate the median values and the bars indicate interquartile range. (B) Gr-1+ cells were depleted from BALB/c mice (n=17) by administration of Gr-1-specific rat monoclonal antibody on days -10, -9 and -8 prior to challenge, as shown on the schema. As a control, additional BALB/c mice (n=9) likewise received an isotyped-matched irrelevant rat antibody. These mice were administered on day -1 prior to challenge 50 μL of the same pooled antiserum (‘L2’, see its titration in Figure 4A) from mice administered three intra-muscular vaccinations at 2-week intervals with 25 μg α11-88x5 multimer formulated with 50 μg aluminum phosphate (n=12 of Gr-1 depleted ‘Gr1’, and n=9 of isotype ‘Iso’ control) or alum only antiserum (‘Alum’ n=5 of the Gr-1 depleted mice). FcγR-deficient mice (−/−, n=7) and control BALB/c mice (+/+, n=12) were likewise each administered 50 μL of the same pooled α11-88x5 antiserum (L2). An additional group (n=10) of control mice received no treatments. All groups of mice were simultaneously challenged with HPV5 PsV and then imaged as in (A). BG=background.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Alam M, Ratner D, 2001. Cutaneous squamous-cell carcinoma. N Engl J Med 344, 975–983. - PubMed
    1. Arroyo Muhr LS, Hultin E, Dillner J, 2021. Transcription of human papillomaviruses in nonmelanoma skin cancers of the immunosuppressed. Int J Cancer 149, 1341–1347. - PubMed
    1. Bernard HU, Burk RD, Chen Z, van Doorslaer K, zur Hausen H, de Villiers EM, 2010. Classification of papillomaviruses (PVs) based on 189 PV types and proposal of taxonomic amendments. Virology 401, 70–79. - PMC - PubMed
    1. Boey L, Curinckx A, Roelants M, Derdelinckx I, Van Wijngaerden E, De Munter P, Vos R, Kuypers D, Van Cleemput J, Vandermeulen C, 2021. Immunogenicity and Safety of the 9-Valent Human Papillomavirus Vaccine in Solid Organ Transplant Recipients and Adults Infected With Human Immunodeficiency Virus (HIV). Clin Infect Dis 73, e661–e671. - PubMed
    1. Bouwes Bavinck JN, Feltkamp MCW, Green AC, Fiocco M, Euvrard S, Harwood CA, Proby CM, Naldi L, Diphoorn JCD, Venturuzzo A, Tessari G, Nindl I, Sampogna F, Abeni D, Neale RE, Goeman JJ, Quint KD, Halk AB, Sneek C, Genders RE, de Koning MNC, Quint WGV, Wieland U, Weissenborn S, Waterboer T, Pawlita M, Pfister H, group E-H-U-C, 2017. Human papillomavirus and posttransplantation cutaneous squamous cell carcinoma: A multicenter, prospective cohort study. Am J Transplant. - PMC - PubMed

Publication types