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Targeting VEGF-A with a vaccine decreases inflammation and joint destruction in experimental arthritis

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Abstract

Objectives

Inflammation and angiogenesis are two tightly linked processes in arthritis, and therapeutic targeting of pro-angiogenic factors may contribute to control joint inflammation and synovitis progression. In this work, we explored whether vaccination against vascular endothelial growth factor (VEGF) ameliorates collagen-induced arthritis (CIA).

Methods

Anti-VEGF vaccines were heterocomplexes consisting of the entire VEGF cytokine (or a VEGF-derived peptide) linked to the carrier protein keyhole limpet hemocyanin (KLH). Two kinds of vaccines were separately tested in two independent experiments of CIA. In the first, we tested a kinoid of the murine cytokine VEGF (VEGF-K), obtained by conjugating VEGF-A to KLH. For the second, we selected two VEGF-A-derived peptide sequences to produce heterocomplexes (Vpep1-K and Vpep2-K). DBA/1 mice were immunized with either VEGF-K, Vpep1-K, or Vpep2-K, before CIA induction. Clinical and histological scores of arthritis, anti-VEGF, anti-Vpep Ab titers, and anti-VEGF Abs neutralizing capacity were determined.

Results

Both VEGF-K and Vpep1-K significantly ameliorated clinical arthritis scores and reduced synovial inflammation and joint destruction at histology. VEGF-K significantly reduced synovial vascularization. None of the vaccines reduced anti-collagen Ab response in mice. Both VEGF-K and Vpep1-K induced persistently high titers of anti-VEGF Abs capable of inhibiting VEGF-A bioactivity.

Conclusion

Vaccination against the pro-angiogenic factor VEGF-A leads to the production of anti-VEGF polyclonal Abs and has a significant anti-inflammatory effect in CIA. Restraining Ab response to a single peptide sequence (Vpep1) with a peptide vaccine effectively protects immunized mice from joint inflammation and destruction.

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Acknowledgments

This research was funded in part by the French Centre National de la Recherche Scientifique and the Laboratory of Excellence Medalis (ANR-10-LABX-0034), Initiative of Excellence (IdEx), Strasbourg University. Pathophysiology, targets, and therapies of rheumatoid arthritis laboratory received grants from University of Paris 13, Agence Nationale de la Recherche (ANR CYTOVAC project) and Inserm (ITMOs IHP and CMN). VEGF kinoid (VEGF-K) was produced by NéoVacs SA (France). We thank Sonia Varela (animal facilities, Paris13 University) and Philippe Haumont for their technical contributions. Special thanks to Delphine Lemeiter, Roxane Hervé, and Matthieu Ribon for their invaluable help with synovial vascularization study. This work is dedicated to the memory of Anna Starzec.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures were approved by the Animal Care and Use Committee of the University of Paris 13 (ethical approval ID: Ce5/2010/036).

Authors’ contributions

L.S. participated in acquisition and interpretation of experimental data, performed statistical analysis, drafted and revised the manuscript. E.D. participated in peptide design, carried out mice immunization, evaluation of clinical arthritis, carried out immunoassay, and helped to draft the manuscript. N.B. carried out mice immunization and evaluation of clinical arthritis. N.M. carried out HUVEC inhibition with sera from peptide immunization. N.S. carried out the synthesis and coupling of peptide vaccines and performed peptide immunoassay. M.M. participated in the design and synthesis of peptide vaccines. G.G.V. conceived a part of the study, participated in the coordination and analysis of the data, and helped to draft the manuscript. EB carried out coupling of VEGF kinoid, immunoassay, and receptor binding inhibition assay. ML participated in the coordination of peptide design and synthesis. H.L. participated in the coordination and analysis of HUVEC inhibition and synovial vascularization. S.M. participated in the design, synthesis and coupling of peptide vaccines, analysis of the data and helped to draft the manuscript. M.C.B. conceived of the study, analysis of the data and helped to draft the manuscript. E.A. conceived of the study, participated in the coordination of the study, analysis of the data and helped to draft the manuscript.

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    Authors and Affiliations

    1. Inserm UMR 1125, Sorbonne Paris Cité - Université Paris 13, 74, rue Marcel Cachin, 93017, Bobigny, France

      Luca Semerano, Emilie Duvallet, Nadia Belmellat, Marie-Christophe Boissier & Eric Assier

    2. Sorbonne Paris Cité - Université Paris 13, 74 rue Marcel Cachin, 93000, Bobigny, France

      Luca Semerano, Emilie Duvallet, Nadia Belmellat, Nicolas Marival, Maëlle Monteil, Marc Lecouvey, Hanna Hlawaty, Marie-Christophe Boissier & Eric Assier

    3. Service de Rhumatologie, Assistance Publique – Hôpitaux de Paris (AP-HP) Groupe hospitalier Avicenne - Jean Verdier – René Muret, 125 rue de Stalingrad, 93000, Bobigny, France

      Luca Semerano & Marie-Christophe Boissier

    4. Inserm UMR 1148, 74 rue Marcel Cachin, 93000, Bobigny, France

      Nicolas Marival & Hanna Hlawaty

    5. CNRS, Immunopathologie et chimie thérapeutique/Laboratory of excellence Medalis, Institut de Biologie Moléculaire et Cellulaire, 15 rue René Descartes, 67000, Strasbourg, France

      Nicolas Schall & Sylviane Muller

    6. CNRS UMR 7244, 74 rue Marcel Cachin, 93000, Bobigny, France

      Maëlle Monteil & Marc Lecouvey

    7. Néovacs SA, 14 Impasse Reille, 75014, Paris, France

      Géraldine Grouard-Vogel & Emilie Bernier

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    1. Luca Semerano
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    Correspondence to Marie-Christophe Boissier.

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    Conflict of interest

    Kinoid technology is patented by NéoVacs SA, and Vpep1 sequence is patented by University of Paris 13. G.G.V. and E.B. are employees of NéoVacs SA. M.C.B. is a member of the scientific board of NéoVacs SA. L.S., E.D., N.B., N.M., N.S., M.M., M.L., H.L., S.M., and E.A. have no competing interests to declare.

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    Luca Semerano and Emilie Duvallet have contributed equally to this work.

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    Semerano, L., Duvallet, E., Belmellat, N. et al. Targeting VEGF-A with a vaccine decreases inflammation and joint destruction in experimental arthritis. Angiogenesis 19, 39–52 (2016). https://doi.org/10.1007/s10456-015-9487-0

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