T cells directed against mutant neo-epitopes drive cancer immunity. However, spontaneous immune recognition of mutations is inefficient. We recently introduced the concept of individualized mutanome vaccines and implemented an RNA-based poly-neo-epitope approach to mobilize immunity against a spectrum of cancer mutations1,2. Here we report the first-in-human application of this concept in melanoma. We set up a process comprising comprehensive identification of individual mutations, computational prediction of neo-epitopes, and design and manufacturing of a vaccine unique for each patient. All patients developed T cell responses against multiple vaccine neo-epitopes at up to high single-digit percentages. Vaccine-induced T cell infiltration and neo-epitope-specific killing of autologous tumour cells were shown in post-vaccination resected metastases from two patients. The cumulative rate of metastatic events was highly significantly reduced after the start of vaccination, resulting in a sustained progression-free survival. Two of the five patients with metastatic disease experienced vaccine-related objective responses. One of these patients had a late relapse owing to outgrowth of β2-microglobulin-deficient melanoma cells as an acquired resistance mechanism. A third patient developed a complete response to vaccination in combination with PD-1 blockade therapy. Our study demonstrates that individual mutations can be exploited, thereby opening a path to personalized immunotherapy for patients with cancer.

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  1. 1.

    et al. Exploiting the mutanome for tumor vaccination. Cancer Res. 72, 1081–1091 (2012)

  2. 2.

    et al. Mutant MHC class II epitopes drive therapeutic immune responses to cancer. Nature 520, 692–696 (2015)

  3. 3.

    et al. Mutational landscape determines sensitivity to PD-1 blockade in non-small cell lung cancer. Science 348, 124–128 (2015)

  4. 4.

    et al. Genetic basis for clinical response to CTLA-4 blockade in melanoma. N. Engl. J. Med. 371, 2189–2199 (2014)

  5. 5.

    et al. Genomic correlates of response to CTLA-4 blockade in metastatic melanoma. Science 350, 207–211 (2015)

  6. 6.

    et al. Clonal neoantigens elicit T cell immunoreactivity and sensitivity to immune checkpoint blockade. Science 351, 1463–1469 (2016)

  7. 7.

    et al. T-cell transfer therapy targeting mutant KRAS in cancer. N. Engl. J. Med. 375, 2255–2262 (2016)

  8. 8.

    et al. High-throughput epitope discovery reveals frequent recognition of neo-antigens by CD4+ T cells in human melanoma. Nat. Med. 21, 81–85 (2015)

  9. 9.

    et al. Cancer exome analysis reveals a T-cell-dependent mechanism of cancer immunoediting. Nature 482, 400–404 (2012)

  10. 10.

    et al. Immunogenicity of somatic mutations in human gastrointestinal cancers. Science 350, 1387–1390 (2015)

  11. 11.

    et al. Predicting immunogenic tumour mutations by combining mass spectrometry and exome sequencing. Nature 515, 572–576 (2014)

  12. 12.

    et al. Checkpoint blockade cancer immunotherapy targets tumour-specific mutant antigens. Nature 515, 577–581 (2014)

  13. 13.

    et al. Cancer immunotherapy. A dendritic cell vaccine increases the breadth and diversity of melanoma neoantigen-specific T cells. Science 348, 803–808 (2015)

  14. 14.

    , & Cancer immunotherapy. Neo approaches to cancer vaccines. Science 348, 760–761 (2015)

  15. 15.

    et al. Intranodal vaccination with naked antigen-encoding RNA elicits potent prophylactic and therapeutic antitumoral immunity. Cancer Res. 70, 9031–9040 (2010)

  16. 16.

    et al. Direct identification of clinically relevant neoepitopes presented on native human melanoma tissue by mass spectrometry. Nat. Commun. 7, 13404 (2016)

  17. 17.

    et al. Mass spectrometry profiling of HLA-associated peptidomes in mono-allelic cells enables more accurate epitope prediction. Immunity 46, 315–326 (2017)

  18. 18.

    & Elements of cancer immunity and the cancer-immune set point. Nature 541, 321–330 (2017)

  19. 19.

    et al. PD-1 blockade induces responses by inhibiting adaptive immune resistance. Nature 515, 568–571 (2014)

  20. 20.

    et al. Pembrolizumab versus ipilimumab in advanced melanoma. N. Engl. J. Med. 372, 2521–2532 (2015)

  21. 21.

    et al. Lack of HLA class I antigen expression by cultured melanoma cells FO-1 due to a defect in B2m gene expression. J. Clin. Invest. 87, 284–292 (1991)

  22. 22.

    et al. Mutations associated with acquired resistance to PD-1 blockade in melanoma. N. Engl. J. Med. 375, 819–829 (2016)

  23. 23.

    et al. Evolving synergistic combinations of targeted immunotherapies to combat cancer. Nat. Rev. Cancer 15, 457–472 (2015)

  24. 24.

    et al. Guidelines for the evaluation of immune therapy activity in solid tumors: immune-related response criteria. Clin. Cancer Res. 15, 7412–7420 (2009)

  25. 25.

    , , , & Generation of tumor-infiltrating lymphocyte cultures for use in adoptive transfer therapy for melanoma patients. J. Immunother. 26, 332–342 (2003)

  26. 26.

    et al. Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. Blood 108, 4009–4017 (2006)

  27. 27.

    & Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25, 1754–1760 (2009)

  28. 28.

    , , & Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol. 10, R25 (2009)

  29. 29.

    , , , & Mapping and quantifying mammalian transcriptomes by RNA-seq. Nat. Methods 5, 621–628 (2008)

  30. 30.

    et al. Immune epitope database analysis resource. Nucleic Acids Res. 40, W525–W530 (2012)

  31. 31.

    et al. Primer3—new capabilities and interfaces. Nucleic Acids Res. 40, e115 (2012)

  32. 32.

    & Enhancements and modifications of primer design program Primer3. Bioinformatics 23, 1289–1291 (2007)

  33. 33.

    BLAT—the BLAST-like alignment tool. Genome Res. 12, 656–664 (2002)

  34. 34.

    et al. Increased antigen presentation efficiency by coupling antigens to MHC class I trafficking signals. J. Immunol. 180, 309–318 (2008)

  35. 35.

    et al. Synthetic mRNAs with superior translation and stability properties. Methods Mol. Biol. 969, 55–72 (2013)

  36. 36.

    et al. Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo. Gene Ther. 17, 961–971 (2010)

  37. 37.

    Magnetic particles for the separation and purification of nucleic acids. Appl. Microbiol. Biotechnol. 73, 495–504 (2006)

  38. 38.

    et al. Functional TCR retrieval from single antigen-specific human T cells reveals multiple novel epitopes. Cancer Immunol. Res. 2, 1230–1244 (2014)

  39. 39.

    , & IMGT/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis. Nucleic Acids Res. 36, W503–W508 (2008)

  40. 40.

    , & Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration. Brief. Bioinform. 14, 178–192 (2013)

  41. 41.

    et al. Optimizing de novo transcriptome assembly from short-read RNA-Seq data: a comparative study. BMC Bioinformatics 12 (Suppl 14), S2 (2011)

  42. 42.

    et al. Luciferase mRNA transfection of antigen presenting cells permits sensitive nonradioactive measurement of cellular and humoral cytotoxicity. J. Immunol. Res. 2016, 9540975 (2016)

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We thank J. de Graaf, I. Eichelbrönner, L. Leppin, L. Giese and S. Vogler, D. Becker, M. Dorner, J. Grützner, M. Hossainzadeh, A. Selmi, S. Wessel, C. Ecker, M. Lochschmitt, B. Schmitz, C. Anft, N. Bidmon, H. Schröder, D. Barea Roldán, C. Walter, S. Wöll, C. Rohde, O. Renz, F. Bayer, C. Kröner, B. Otte, T. Stricker, M. Drude S. Petri, M. Mechler, L. Hebich, B. Steege, A. Oelbermann, J. Schwarz, C. Britten, J. C. Castle and B. Pless for technical support, project management and advice. We thank A. Tüttenberg for support with a figure. We thank I. Mellman, L. Delamarre and G. Fine for critical reading of the manuscript. We thank K. Sahin for her advice. The study was supported by the CI3 cluster program of the Federal Ministry of Education and Research (BMBF).

Author information

Author notes

    • Carmen Loquai
    •  & Özlem Türeci

    These authors contributed equally to this work.


  1. Biopharmaceutical New Technologies (BioNTech) Corporation, An der Goldgrube 12, 55131 Mainz, Germany

    • Ugur Sahin
    • , Evelyna Derhovanessian
    • , Matthias Miller
    • , Björn-Philipp Kloke
    • , Petra Simon
    • , Valesca Bukur
    • , Ulrich Luxemburger
    • , Tana Omokoko
    • , Mathias Vormehr
    • , Anna Paruzynski
    • , Andreas N. Kuhn
    • , Janina Buck
    • , Sandra Heesch
    • , Katharina H. Schreeb
    • , Felicitas Müller
    • , Inga Ortseifer
    • , Isabel Vogler
    • , Eva Godehardt
    • , Andrea Breitkreuz
    • , Claudia Tolliver
    • , Jan Diekmann
    • , Alexandra-Kemmer Brück
    • , Meike Witt
    • , Martina Zillgen
    • , David Langer
    • , Stefanie Bolte
    • , Mustafa Diken
    • , Sebastian Kreiter
    •  & Christoph Huber
  2. TRON – Translational Oncology at the University Medical Center of Johannes Gutenberg University gGmbH, Freiligrathstraße 12, 55131 Mainz, Germany

    • Ugur Sahin
    • , Martin Löwer
    • , Valesca Bukur
    • , Arbel D. Tadmor
    • , Barbara Schrörs
    • , Christian Albrecht
    • , Sebastian Attig
    • , Richard Rae
    • , Martin Suchan
    • , Goran Martic
    • , Patrick Sorn
    • , Andree Rothermel
    • , Barbara Kasemann
    • , Mustafa Diken
    • , Sebastian Kreiter
    •  & Christoph Huber
  3. University Medical Center of the Johannes Gutenberg University, Langenbeckstraße 1, 55131 Mainz, Germany

    • Ugur Sahin
    • , Mathias Vormehr
    • , Sebastian Attig
    • , Alexander Hohberger
    • , Stephan Grabbe
    • , Christoph Huber
    •  & Carmen Loquai
  4. EUFETS GmbH, Vollmersbachstraße 66, 55743 Idar-Oberstein, Germany

    • Janko Ciesla
    •  & Olga Waksmann
  5. Medical University of Vienna, Spitalgasse 23, 1090 Vienna, Austria

    • Romina Nemecek
    •  & Christoph Höller
  6. German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany

    • Christoffer Gebhardt
    •  & Jochen Utikal
  7. University Medical Center Mannheim, Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68135 Mannheim, Germany

    • Christoffer Gebhardt
    •  & Jochen Utikal
  8. CI3 - Cluster for Individualized Immunointervention e.V, Hölderlinstraße 8, 55131 Mainz, Germany

    • Özlem Türeci


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U.S. conceptualized the work and strategy. E.D., Pe.Si., T.O., I.O., I.V., S.A., A.R. and B.K. planned and analysed experiments. E.G., R.R., A.B., C.T. and A.H. did experiments. M.L., V.B., A.D.T., B.S., C.A., A.P. and Pa.So. performed and analysed NGS runs. A.N.K., J.B. and J.C. manufactured the RNA vaccines, O.W., M.W. and M.Z. performed quality assurance. M.M., B.K., S.H., K.H.S., F.M., A.K.-B., D.L. and S.B. managed sample logistics. R.N., C.G., S.G., C.H., J.U. are clinical investigators. C.L. is the principal clinical investigator. U.L., J.D., M.D. and S.K. supported clinical grade assays. U.S., ÖT supported by E.D., M.V., Pe.Si., M.L, M.M., B.S. interpreted data and wrote the manuscript. All authors supported the revision of the manuscript.

Competing interests

Some of the authors are employees at BioNTech AG (Mainz, Germany) as mentioned in the affiliations. U.S. is stock owner of BioNTech AG (Mainz, Germany). U.S., M.L., B.S., M.V., A.N.K., M.D., A.T., Ö.T. and S.K. are inventors on patents and patent applications, which cover parts of this article.

Corresponding author

Correspondence to Ugur Sahin.

Reviewer Information Nature thanks C. Melief and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Supplementary information

Excel files

  1. 1.

    Supplementary Table 1

    This table lists the neo-epitope vaccine sequence for patient P04.

  2. 2.

    Supplementary Table 2

    This table summarizes all identified neo-epitopes across patients.

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