Barley produced Culicoides allergens are suitable for monitoring the immune response of horses immunized with E. coli expressed allergens.

Jonsdottir, Sigridur; Stefansdottir, Sara Bjork; Kristinarson, Sæmundur Bjarni; Svansson, Vilhjalmur; Bjornsson, Jon Mar; Runarsdottir, Arna; Wagner, Bettina; Marti, Eliane Isabelle; Torsteinsdottir, Sigurbjorg (2018). Barley produced Culicoides allergens are suitable for monitoring the immune response of horses immunized with E. coli expressed allergens. Veterinary immunology and immunopathology, 201, pp. 32-37. Elsevier 10.1016/j.vetimm.2018.05.005

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Insect bite hypersensitivity is an allergic dermatitis of horses caused by bites of Culicoides midges. Sufficient amount of pure, endotoxin-free allergens is a prerequisite for development and monitoring of preventive and therapeutic allergen immunotherapy. Aims of the study were to compare the Culicoides nubeculosus (Cul n) allergens Cul n 3 and Cul n 4, produced in transgenic barley grains with the corresponding E. coli or insect cells expressed proteins for measuring antibody and cytokine responses. Allergen-specific IgG responses were measured by ELISA in sera from twelve horses not exposed to Culicoides, before and after vaccination with E. coli-rCul n 3 and 4. Before vaccination no IgG binding to the barley and insect cell produced proteins was detected and a similar increase in specific IgG was observed after vaccination. While IgG levels to the E.coli expressed proteins were higher in the post-vaccination sera, some background binding was observed pre-vaccination. In vitro re-stimulation of PBMC was performed for measurements of cytokines. E. coli expressed proteins resulted in high background in PBMC from non-vaccinated controls. The barley and insect cell expressed proteins induced similar amount of IFN-γ and IL-4 in PBMC from vaccinated horses. Barley produced allergens are promising tools for use in immunoassays.

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