Cancer Immunology, Immunotherapy

, Volume 66, Issue 9, pp 1163–1173 | Cite as

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

  • Sanne Samuels
  • A. Marijne Heeren
  • Henry J. M. A. A. Zijlmans
  • Marij J. P. Welters
  • Joost H. van den Berg
  • Daisy Philips
  • Pia Kvistborg
  • Ilina Ehsan
  • Suzy M. E. Scholl
  • Bastiaan Nuijen
  • Ton N. M. Schumacher
  • Marc van Beurden
  • Ekaterina S. Jordanova
  • John B. A. G. Haanen
  • Sjoerd H. van der Burg
  • Gemma G. KenterEmail author
Original Article



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).


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.


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.


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.


VIN HPV Immunotherapy DNA vaccine Immunogenicity Safety 



Common terminology criteria for adverse events


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


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


Synthetic long peptide


Tetanus toxin fragment C


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


Usual type vulvar intraepithelial neoplasia


Vulvar intraepithelial neoplasia



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 ( 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).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

262_2017_2006_MOESM1_ESM.pdf (6.1 mb)
Supplementary material 1 (PDF 6268 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Sanne Samuels
    • 1
  • A. Marijne Heeren
    • 2
  • Henry J. M. A. A. Zijlmans
    • 1
  • Marij J. P. Welters
    • 3
  • Joost H. van den Berg
    • 4
  • Daisy Philips
    • 5
  • Pia Kvistborg
    • 5
  • Ilina Ehsan
    • 3
  • Suzy M. E. Scholl
    • 6
  • Bastiaan Nuijen
    • 7
  • Ton N. M. Schumacher
    • 5
  • Marc van Beurden
    • 1
  • Ekaterina S. Jordanova
    • 1
  • John B. A. G. Haanen
    • 5
    • 8
  • Sjoerd H. van der Burg
    • 3
  • Gemma G. Kenter
    • 1
    Email author
  1. 1.Department of GynecologyCenter for Gynecologic Oncology AmsterdamAmsterdamThe Netherlands
  2. 2.Department of Medical OncologyVU University Medical Center-Cancer Center AmsterdamAmsterdamThe Netherlands
  3. 3.Department of Clinical OncologyLeiden University Medical CenterLeidenThe Netherlands
  4. 4.Amsterdam Biotherapeutics Unit (AmBTU)AmsterdamThe Netherlands
  5. 5.Department of ImmunologyNetherlands Cancer Institute, Antoni van LeeuwenhoekAmsterdamThe Netherlands
  6. 6.Department of Medical OncologyInstitut CurieParisFrance
  7. 7.Department of Pharmacy and PharmacologyNetherlands Cancer Institute, Antoni van LeeuwenhoekAmsterdamThe Netherlands
  8. 8.Department of Medical OncologyNetherlands Cancer Institute, Antoni van LeeuwenhoekAmsterdamThe Netherlands

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