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Immune reactivity to type VII collagen: implications for gene therapy of recessive dystrophic epidermolysis bullosa

Gene Therapy volume 17, pages 930–937 (2010)Cite this article

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Abstract

Recessive dystrophic epidermolysis bullosa (RDEB) is a severe genodermatosis caused by loss-of-function mutations in COL7A1 encoding type VII collagen, the component of anchoring fibrils. As exogenous type VII collagen may elicit a deleterious immune response in RDEB patients during upcoming clinical trials of gene therapies or protein replacement therapies, we developed enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunosorbent spot (ELISPOT) assays to analyze B- and T-cell responses, to the full-length type VII collagen. The ELISA was highly sensitive and specific when tested against sera from 41 patients with epidermolysis bullosa acquisita (EBA), and the IFN-γ ELISPOT detected a cellular response that correlated with ongoing EBA manifestations. Both tests were next applied to assess the risk of an immune response to type VII collagen in seven RDEB patients with a range of type VII collagen expression profiles. Immune responses against type VII collagen were dependent on the expression of type VII collagen protein, and consequently on the nature and position of the respective COL7A1 mutations. These immunologic tests will be helpful for the selection of RDEB patients for future clinical trials aiming at restoring type VII collagen expression, and in monitoring their immune response to type VII collagen after treatment.

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Acknowledgements

We thank Dr Mei Chen for the gift of the polyclonal anti-NC1 antibody, and Professor Yann Barrandon for sharing the immortalized RDEB keratinocyte cell line and for the protein extract of patient 10 cells. We are grateful to Drs Joost van Meerwijk and Paola Romagnoli for fruitful discussion, and Drs Gilbert Fournié and Abdelhadi Saoudi for their critical reading of the manuscript. This work was supported by the Epidermolyse Bulleuse Association d'Entraide (EBAE), the Association Française contre les Myopathies (AFM), the Geneskin European coordination action project, the Therapeuskin European specific targeted research project, the Midi-Pyrénées Region, the French Ministry of Health through the reference centers for genetic skin diseases, and the Agence Nationale pour la Recherche (ANR, DEBCURE, No. ANR-07-BLAN-0105).

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Authors and Affiliations

  1. Inserm, U563, Toulouse, France

    V Pendaries, G Gasc, M Titeux, J E Mejía, A Décha & A Hovnanian

  2. Université Toulouse III Paul Sabatier, Toulouse, France

    V Pendaries, G Gasc, M Titeux, J E Mejía & A Décha

  3. Département d'Immunologie et Histocompatibilité, Hôpital Saint-Louis, Paris, France

    C Leroux & P Loiseau

  4. Ecole Nationale Supérieure Agronomique de Toulouse, Auzeville-Tolosane

    Z G Vitezica

  5. Centre de Référence sur les Maladies Génétiques à Expression Cutanée (MAGEC), Hôpital Necker-Enfants Malades, Paris, France

    C Bodemer

  6. Département de Dermatologie, Hôpital Necker-Enfants malades, Paris, France

    C Bodemer & A Hovnanian

  7. Université Paris V René Descartes, Paris, France

    C Bodemer & A Hovnanian

  8. Centre de Référence des Maladies Bulleuses Auto-immunes Ile de France, Hôpital Avicenne, Bobigny, France

    C Prost-Squarcioni

  9. UFR Léonard de Vinci, Laboratoire d'Histologie et de Thérapie Génique, Université Paris 13, Bobigny, France

    C Prost-Squarcioni

  10. Département de Génétique – Hôpital Necker-Enfants malades, Paris, France

    A Hovnanian

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  1. V Pendaries
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Pendaries, V., Gasc, G., Titeux, M. et al. Immune reactivity to type VII collagen: implications for gene therapy of recessive dystrophic epidermolysis bullosa. Gene Ther 17, 930–937 (2010). https://doi.org/10.1038/gt.2010.36

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