Bases structurales de la régulation des cytokines par les héparanes sulfates : régulation génique et optimisation d’un inhibiteur de l’interféron-gamma.

Abstract : Interferon-γ (IFNγ) is a strong immunomodulating cytokine with some antiviral activity. It has two ligands for which it has high affinities: a receptor through which it transmits its signals, and complex polysaccharides of the heparan sulphate (HS) family. Both are situated on the cellular surface. In vivo, binding on the HS permits local concentration of the cytokine, and regulates its biological activity via a partial protection of the C-terminal region. This C-terminal region, characterised by two basic domains, D1 and D2, is implicated in the recognition of the receptor and the HS. In this context, we investigated the structural features for the interaction of IFNγ, and more specifically the importance of his C-terminus, with both of his cellular ligands. Therefore, we produced, purified and examined various mutants of IFNγ, including points, multiples and deletions mutants. There ability to bind to HS and the IFNγ receptor is examined by SPR and there influence on IFNγ's antiviral activity is determined. The thermodynamics complexation of IFNγ with the HS-oligosaccharides is examined. Moreover, we have prepared an oligosaccharidic library derived from HS. By screening this library for its capacity to bind IFNγ, we have demonstrated that the cytokine recognizes a particular sulphated pattern. Finely, we tried to crystallize the IFNγ:HS-oligosaccharide complex, without obtaining diffracting crystals yet. These studies contribute to clarify the mechanism of recognition of IFNγ by the HS. This would enable us to design a mimic of the interaction site for IFNγ on the HS, who inhibits inappropriate signaling of the cytokine. Finely, a detailed comprehension of the interaction of IFNγ with his receptor and with the HS needs to be established to fully understand how IFNγ migrates for the HS to its receptor.
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Els Saesen. Bases structurales de la régulation des cytokines par les héparanes sulfates : régulation génique et optimisation d’un inhibiteur de l’interféron-gamma.. Sciences agricoles. Université de Grenoble, 2013. Français. ⟨NNT : 2013GRENV004⟩. ⟨tel-00949161⟩

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