Skip to main content

Advertisement

SpringerLink
Log in

HPV16 E7 DNA tattooing: safety, immunogenicity, and clinical response in patients with HPV-positive vulvar intraepithelial neoplasia

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Background

Usual type vulvar intraepithelial neoplasia (uVIN) is caused by HPV, predominantly type 16. Several forms of HPV immunotherapy have been studied, however, clinical results could be improved. A novel intradermal administration route, termed DNA tattooing, is superior in animal models, and was tested for the first time in humans with a HPV16 E7 DNA vaccine (TTFC-E7SH).

Methods

The trial was designed to test safety, immunogenicity, and clinical response of TTFC-E7SH in twelve HPV16+ uVIN patients. Patients received six vaccinations via DNA tattooing. The first six patients received 0.2 mg TTFC-E7SH and the next six 2 mg TTFC-E7SH. Vaccine-specific T-cell immunity was evaluated by IFNγ-ELISPOT and multiparametric flow cytometry.

Results

Only grade I-II adverse events were observed upon TTFC-E7SH vaccination. The ELISPOT analysis showed in 4/12 patients a response to the peptide pool containing shuffled E7 peptides. Multiparametric flow cytometry showed low CD4+ and/or CD8+ T-cell responses as measured by increased expression of PD-1 (4/12 in both), CTLA-4 (2/12 and 3/12), CD107a (5/12 and 4/12), or the production of IFNγ (2/12 and 1/12), IL-2 (3/12 and 4/12), TNFα (2/12 and 1/12), and MIP1β (3/12 and 6/12). At 3 months follow-up, no clinical response was observed in any of the twelve vaccinated patients.

Conclusion

DNA tattoo vaccination was shown to be safe. A low vaccine-induced immune response and no clinical response were observed in uVIN patients after TTFC-E7SH DNA tattoo vaccination. Therefore, a new phase I/II trial with an improved DNA vaccine format is currently in development for patients with uVIN.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

CTCAE:

Common terminology criteria for adverse events

sig-HELP-E6SH-kdel:

Vaccine encoding the fusion protein of the carrier sequence sig-HELP-kdel and the shuffled version of the HPV16 E6 oncoprotein

sig-HELP-E7SH-kdel:

Vaccine encoding the fusion protein of the carrier sequence sig-HELP-kdel and the shuffled version of the HPV16 E7 oncoprotein

SLP:

Synthetic long peptide

TTFC:

Tetanus toxin fragment C

TTFC-E7SH:

Vaccine encoding the fusion protein of TTFC and a shuffled variant of HPV16 E7

uVIN:

Usual type vulvar intraepithelial neoplasia

VIN:

Vulvar intraepithelial neoplasia

References

  1. Sideri M, Jones RW, Wilkinson EJ, Preti M, Heller DS, Scurry J, Haefner H, Neill S (2005) Squamous vulvar intraepithelial neoplasia: 2004 modified terminology, ISSVD Vulvar Oncology Subcommittee. J Reprod Med 50(11):807–810

    PubMed  Google Scholar 

  2. Buscema J, Naghashfar Z, Sawada E, Daniel R, Woodruff JD, Shah K (1988) The predominance of human papillomavirus type 16 in vulvar neoplasia. Obstet Gynecol 71(4):601–606

    CAS  PubMed  Google Scholar 

  3. Hording U, Junge J, Poulsen H, Lundvall F (1995) Vulvar intraepithelial neoplasia III: a viral disease of undetermined progressive potential. Gynecol Oncol 56(2):276–279

    Article  CAS  PubMed  Google Scholar 

  4. Serrano B, de Sanjose S, Tous S, Quiros B, Munoz N, Bosch X, Alemany L (2015) Human papillomavirus genotype attribution for HPVs 6, 11, 16, 18, 31, 33, 45, 52 and 58 in female anogenital lesions. Eur J Cancer 51(13):1732–1741

    Article  PubMed  Google Scholar 

  5. Smith JS, Backes DM, Hoots BE, Kurman RJ, Pimenta JM (2009) Human papillomavirus type-distribution in vulvar and vaginal cancers and their associated precursors. Obstet Gynecol 113(4):917–924

    Article  PubMed  Google Scholar 

  6. van Beurden M, ten Kate FJ, Smits HL, Berkhout RJ, de Craen AJ, van der Vange N, Lammes FB, ter Schegget J (1995) Multifocal vulvar intraepithelial neoplasia grade III and multicentric lower genital tract neoplasia is associated with transcriptionally active human papillomavirus. Cancer 75(12):2879–2884

    Article  PubMed  Google Scholar 

  7. Kuhn L, Sun XW, Wright TC Jr (1999) Human immunodeficiency virus infection and female lower genital tract malignancy. Curr Opin Obstet Gynecol 11(1):35–39

    Article  CAS  PubMed  Google Scholar 

  8. van de Nieuwenhof HP, van der Avoort IA, de Hullu JA (2008) Review of squamous premalignant vulvar lesions. Crit Rev Oncol Hematol 68(2):131–156

    Article  PubMed  Google Scholar 

  9. Jones RW, Rowan DM, Stewart AW (2005) Vulvar intraepithelial neoplasia: aspects of the natural history and outcome in 405 women. Obstet Gynecol 106(6):1319–1326

    Article  PubMed  Google Scholar 

  10. van Seters M, van Beurden M, de Craen AJ (2005) Is the assumed natural history of vulvar intraepithelial neoplasia III based on enough evidence? A systematic review of 3322 published patients. Gynecol Oncol 97(2):645–651

    Article  PubMed  Google Scholar 

  11. Andreasson B, Bock JE (1985) Intraepithelial neoplasia in the vulvar region. Gynecol Oncol 21(3):300–305

    Article  CAS  PubMed  Google Scholar 

  12. Rettenmaier MA, Berman ML, DiSaia PJ (1987) Skinning vulvectomy for the treatment of multifocal vulvar intraepithelial neoplasia. Obstet Gynecol 69(2):247–250

    CAS  PubMed  Google Scholar 

  13. Sykes P, Smith N, McCormick P, Frizelle FA (2002) High-grade vulval intraepithelial neoplasia (VIN 3): a retrospective analysis of patient characteristics, management, outcome and relationship to squamous cell carcinoma of the vulva 1989–1999. Aust N Z J Obstet Gynaecol 42(1):69–74

    Article  PubMed  Google Scholar 

  14. Daayana S, Elkord E, Winters U, Pawlita M, Roden R, Stern PL, Kitchener HC (2010) Phase II trial of imiquimod and HPV therapeutic vaccination in patients with vulval intraepithelial neoplasia. Br J Cancer 102(7):1129–1136

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kenter GG, Welters MJ, Valentijn AR, Lowik MJ, Berends-van der Meer DM, Vloon AP, Essahsah F, Fathers LM, Offringa R, Drijfhout JW, Wafelman AR, Oostendorp J, Fleuren GJ, van der Burg SH, Melief CJ (2009) Vaccination against HPV-16 oncoproteins for vulvar intraepithelial neoplasia. N Engl J Med 361(19):1838–1847

    Article  CAS  PubMed  Google Scholar 

  16. van Poelgeest MI, Welters MJ, Vermeij R, Stynenbosch LF, Loof NM, Berends-van der Meer DM, Lowik MJ, Hamming IL, van Esch EM, Hellebrekers BW, van Beurden M, Schreuder HW, Kagie MJ, Trimbos JB, Fathers LM, Daemen T, Hollema H, Valentijn AR, Oostendorp J, Oude Elberink JH, Fleuren GJ, Bosse T, Kenter GG, Stijnen T, Nijman HW, Melief CJ, van der Burg SH (2016) Vaccination against Oncoproteins of HPV16 for noninvasive vulvar/vaginal lesions: lesion clearance is related to the strength of the T-cell response. Clin Cancer Res 22(10):2342–2350

    Article  PubMed  Google Scholar 

  17. Welters MJ, Kenter GG, de Vos van Steenwijk PJ, Lowik MJ, Berends-van der Meer DM, Essahsah F, Stynenbosch LF, Vloon AP, Ramwadhdoebe TH, Piersma SJ, van der Hulst JM, Valentijn AR, Fathers LM, Drijfhout JW, Franken KL, Oostendorp J, Fleuren GJ, Melief CJ, van der Burg SH (2010) Success or failure of vaccination for HPV16-positive vulvar lesions correlates with kinetics and phenotype of induced T-cell responses. Proc Natl Acad Sci USA 107 (26):11895–11899

  18. Trimble CL, Morrow MP, Kraynyak KA, Shen X, Dallas M, Yan J, Edwards L, Parker RL, Denny L, Giffear M, Brown AS, Marcozzi-Pierce K, Shah D, Slager AM, Sylvester AJ, Khan A, Broderick KE, Juba RJ, Herring TA, Boyer J, Lee J, Sardesai NY, Weiner DB, Bagarazzi ML (2015) Safety, efficacy, and immunogenicity of VGX-3100, a therapeutic synthetic DNA vaccine targeting human papillomavirus 16 and 18 E6 and E7 proteins for cervical intraepithelial neoplasia 2/3: a randomised, double-blind, placebo-controlled phase 2b trial. Lancet 386(10008):2078–2088

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Rice J, Ottensmeier CH, Stevenson FK (2008) DNA vaccines: precision tools for activating effective immunity against cancer. Nat Rev Cancer 8(2):108–120

    Article  CAS  PubMed  Google Scholar 

  20. Bins AD, Jorritsma A, Wolkers MC, Hung CF, Wu TC, Schumacher TN, Haanen JB (2005) A rapid and potent DNA vaccination strategy defined by in vivo monitoring of antigen expression. Nat Med 11(8):899–904

    Article  CAS  PubMed  Google Scholar 

  21. Verstrepen BE, Bins AD, Rollier CS, Mooij P, Koopman G, Sheppard NC, Sattentau Q, Wagner R, Wolf H, Schumacher TN, Heeney JL, Haanen JB (2008) Improved HIV-1 specific T-cell responses by short-interval DNA tattooing as compared to intramuscular immunization in non-human primates. Vaccine 26(26):3346–3351

    Article  CAS  PubMed  Google Scholar 

  22. van den Berg JH, Nujien B, Beijnen JH, Vincent A, van Tinteren H, Kluge J, Woerdeman LA, Hennink WE, Storm G, Schumacher TN, Haanen JB (2009) Optimization of intradermal vaccination by DNA tattooing in human skin. Hum Gene Ther 20(3):181–189. doi:10.1089/hgt.2008.073

    Article  PubMed  PubMed Central  Google Scholar 

  23. Oosterhuis K, Ohlschlager P, van den Berg JH, Toebes M, Gomez R, Schumacher TN, Haanen JB (2011) Preclinical development of highly effective and safe DNA vaccines directed against HPV 16 E6 and E7. Int J Cancer 129(2):397–406

    Article  CAS  PubMed  Google Scholar 

  24. Henken FE, Oosterhuis K, Ohlschlager P, Bosch L, Hooijberg E, Haanen JB, Steenbergen RD (2012) Preclinical safety evaluation of DNA vaccines encoding modified HPV16 E6 and E7. Vaccine 30(28):4259–4266

    Article  CAS  PubMed  Google Scholar 

  25. Trimble CL, Peng S, Kos F, Gravitt P, Viscidi R, Sugar E, Pardoll D, Wu TC (2009) A phase I trial of a human papillomavirus DNA vaccine for HPV16+ cervical intraepithelial neoplasia 2/3. Clin Cancer Res 15(1):361–367

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Przybylowski M, Bartido S, Borquez-Ojeda O, Sadelain M, Riviere I (2007) Production of clinical-grade plasmid DNA for human Phase I clinical trials and large animal clinical studies. Vaccine 25(27):5013–5024

    Article  CAS  PubMed  Google Scholar 

  27. Quaak SG, van den Berg JH, Toebes M, Schumacher TN, Haanen JB, Beijnen JH, Nuijen B (2008) GMP production of pDERMATT for vaccination against melanoma in a phase I clinical trial. Eur J Pharm Biopharm 70(2):429–438

    Article  CAS  PubMed  Google Scholar 

  28. Urthaler J, Buchinger W, Necina R (2005) Improved downstream process for the production of plasmid DNA for gene therapy. Acta Biochim Pol 52(3):703–711

    CAS  PubMed  Google Scholar 

  29. van Poelgeest MI, Welters MJ, van Esch EM, Stynenbosch LF, Kerpershoek G, van Persijn van Meerten EL, van den Hende M, Lowik MJ, Berends-van der Meer DM, Fathers LM, Valentijn AR, Oostendorp J, Fleuren GJ, Melief CJ, Kenter GG, van der Burg SH (2013) HPV16 synthetic long peptide (HPV16-SLP) vaccination therapy of patients with advanced or recurrent HPV16-induced gynecological carcinoma, a phase II trial. J Transl Med 11:88

  30. Oosterhuis K, Aleyd E, Vrijland K, Schumacher TN, Haanen JB (2012) Rational design of DNA vaccines for the induction of human papillomavirus type 16 E6- and E7-specific cytotoxic T-cell responses. Hum Gene Ther 23(12):1301–1312

    Article  CAS  PubMed  Google Scholar 

  31. Gros A, Parkhurst MR, Tran E, Pasetto A, Robbins PF, Ilyas S, Prickett TD, Gartner JJ, Crystal JS, Roberts IM, Trebska-McGowan K, Wunderlich JR, Yang JC, Rosenberg SA (2016) Prospective identification of neoantigen-specific lymphocytes in the peripheral blood of melanoma patients. Nat Med 22(4):433–438

    Article  CAS  PubMed  Google Scholar 

  32. Ahmadzadeh M, Johnson LA, Heemskerk B, Wunderlich JR, Dudley ME, White DE, Rosenberg SA (2009) Tumor antigen-specific CD8 T cells infiltrating the tumor express high levels of PD-1 and are functionally impaired. Blood 114(8):1537–1544

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Hokey DA, Johnson FB, Smith J, Weber JL, Yan J, Hirao L, Boyer JD, Lewis MG, Makedonas G, Betts MR, Weiner DB (2008) Activation drives PD-1 expression during vaccine-specific proliferation and following lentiviral infection in macaques. Eur J Immunol 38(5):1435–1445

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Fourcade J, Sun Z, Benallaoua M, Guillaume P, Luescher IF, Sander C, Kirkwood JM, Kuchroo V, Zarour HM (2010) Upregulation of Tim-3 and PD-1 expression is associated with tumor antigen-specific CD8+ T cell dysfunction in melanoma patients. J Exp Med 207(10):2175–2186

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. van der Burg SH, Piersma SJ, de Jong A, van der Hulst JM, Kwappenberg KM, van den Hende M, Welters MJ, Van Rood JJ, Fleuren GJ, Melief CJ, Kenter GG, Offringa R (2007) Association of cervical cancer with the presence of CD4+ regulatory T cells specific for human papillomavirus antigens. Proc Natl Acad Sci USA 104(29):12087–12092

    Article  PubMed  PubMed Central  Google Scholar 

  36. Santegoets SJ, Dijkgraaf EM, Battaglia A, Beckhove P, Britten CM, Gallimore A, Godkin A, Gouttefangeas C, de Gruijl TD, Koenen HJ, Scheffold A, Shevach EM, Staats J, Tasken K, Whiteside TL, Kroep JR, Welters MJ, van der Burg SH (2015) Monitoring regulatory T cells in clinical samples: consensus on an essential marker set and gating strategy for regulatory T cell analysis by flow cytometry. Cancer Immunol Immunother 64(10):1271–1286

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Dr. Freek Groenman of the Netherlands Cancer Institute – Antoni van Leeuwenhoek hospital (NKI-AVL, Amsterdam, the Netherlands) for including patients. We also like to thank the Rational molecular Assessment Innovative Drug selection (RAIDs) consortium (http://www.raids-fp7.eu). This trial is part of the RAIDs project and received funding from the European Union’s Seventh Program for Research, Technological Development, and Demonstration (Grant No. 304810).

Author information

Authors and Affiliations

  1. Department of Gynecology, Center for Gynecologic Oncology Amsterdam, P.O. Box 90203, 1006 BE, Amsterdam, The Netherlands

    Sanne Samuels, Henry J. M. A. A. Zijlmans, Marc van Beurden, Ekaterina S. Jordanova & Gemma G. Kenter

  2. Department of Medical Oncology, VU University Medical Center-Cancer Center Amsterdam, De Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands

    A. Marijne Heeren

  3. Department of Clinical Oncology, Leiden University Medical Center, P.O. Box 9600, 2300 RC, Leiden, The Netherlands

    Marij J. P. Welters, Ilina Ehsan & Sjoerd H. van der Burg

  4. Amsterdam Biotherapeutics Unit (AmBTU), Louwesweg 6, 1066 EC, Amsterdam, The Netherlands

    Joost H. van den Berg

  5. Department of Immunology, Netherlands Cancer Institute, Antoni van Leeuwenhoek, P.O. Box 90203, 1006 BE, Amsterdam, The Netherlands

    Daisy Philips, Pia Kvistborg, Ton N. M. Schumacher & John B. A. G. Haanen

  6. Department of Medical Oncology, Institut Curie, 25 Rue d’Ulm, 75005, Paris, France

    Suzy M. E. Scholl

  7. Department of Pharmacy and Pharmacology, Netherlands Cancer Institute, Antoni van Leeuwenhoek, P.O. Box 90203, 1006 BE, Amsterdam, The Netherlands

    Bastiaan Nuijen

  8. Department of Medical Oncology, Netherlands Cancer Institute, Antoni van Leeuwenhoek, P.O. Box 90203, 1006 BE, Amsterdam, The Netherlands

    John B. A. G. Haanen

Authors
  1. Sanne Samuels
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Marijne Heeren
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Henry J. M. A. A. Zijlmans
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marij J. P. Welters
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Joost H. van den Berg
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daisy Philips
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Pia Kvistborg
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ilina Ehsan
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Suzy M. E. Scholl
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Bastiaan Nuijen
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Ton N. M. Schumacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Marc van Beurden
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Ekaterina S. Jordanova
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. John B. A. G. Haanen
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Sjoerd H. van der Burg
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Gemma G. Kenter
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gemma G. Kenter.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 6268 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Samuels, S., Marijne Heeren, A., Zijlmans, H.J.M.A.A. et al. HPV16 E7 DNA tattooing: safety, immunogenicity, and clinical response in patients with HPV-positive vulvar intraepithelial neoplasia. Cancer Immunol Immunother 66, 1163–1173 (2017). https://doi.org/10.1007/s00262-017-2006-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-017-2006-y

Keywords

Search

Navigation