Modification des dodécaèdres bases de l'adénovirus de sérotype 3 : design et caractérisation d'un nouveau vecteur multi-épitopique polyvalent

Abstract : Some human adenoviruses (HAdV) such as adenovirus derived from serotype 3 (belonging to subgroup B) are able to form virus-like particles composed of the two proteins involved in viral entry: the penton base and the fiber (= penton). Indeed, 12 pentons are able to self-assemble in a symmetrical manner to form penton dodecahedron (PtDd). In the present work, we modified and characterized the base dodecahedron (BsDd = PtDd without fiber) of HAdV3 in order to create a versatile multi-epitopic platform named ADDomer (ADenovirus Dodecamer). We have created a genetic platform allowing easy insertion of epitope(s) of interest (s) thanks to synthetic biology. The insertion of sequences encoding a peptide of interest in the ADDomer gene enable a multivalent exposure at the surface of the VLP due to the pentamerization then to the dodecamerization of the penton base. ADDomer has been produced and characterized to assess its ability to vectorize linear or structurally complex epitopes. We then designed a second vectorization strategy, still based on the ADDomer, but using the interaction penton base / fibre. A peptide mimicking the part of the Ad3 fiber interacting with the penton base (the 20 N-terminal residues) has been designed to serve as an adaptor forming covalent bonds with the ADDomer.The behavior of the ADDomer in vivo has been studied in a vaccine context. For this, we injected the ADDomer in mice to validate its transport to the lymphatic system. We have also demonstrated that ADDomer is able to internalize monocytes and dendritic cells derived from monocytes (MoDC) and induces the specific characters of MoDC maturation. Based on these results, we generated an ADDomer vectorizing an epitope of the Chikungunya virus (ADDomer TevChik) described to be the target of neutralizing antibodies of patients who have been infected by this virus. To conclude this in vivo study, we assessed the ability of ADDomer TevChik to induce the anti-epitopic response and thus demonstrated that the way the epitope is displayed on the surface of the ADDomer was important to obtain a meaningful response.
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Charles Vragniau. Modification des dodécaèdres bases de l'adénovirus de sérotype 3 : design et caractérisation d'un nouveau vecteur multi-épitopique polyvalent. Biologie structurale [q-bio.BM]. Université Grenoble Alpes, 2018. Français. ⟨NNT : 2018GREAV061⟩. ⟨tel-02110396⟩

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