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Immunol."],"abstract":"Introduction<\/jats:title>Insect bite hypersensitivity (IBH) is an IgE-mediated dermatitis in horses caused by bites of Culicoides<\/jats:italic> spp. The allergens are salivary gland proteins from these insects, and nine major allergens from Culicoides obsoletus<\/jats:italic> have been identified and expressed in E. coli<\/jats:italic>. However, proteins expressed in procaryotic systems have limitations in cellular assays, particularly in functional assays assessing the allergen-induced release of mediators in vitro<\/jats:italic>, such as sulphidoleukotrienes (sLT) from basophils. The aims of the study were to produce functional Culicoides<\/jats:italic> allergens in insect cells, to assess their allergenicity using a sLT release assay, and to relate the sLT release with IgE sensitization to the respective allergens using ELISA.<\/jats:p><\/jats:sec>Methods<\/jats:title>Eight major Culicoides obsoletus<\/jats:italic> allergens (Cul o 1P, Cul o 2P, Cul o 3, Cul o 5, Cul o 7, Cul o 8, Cul o 9, and Cul o 11) were expressed in insect cells and purified. sLT release from peripheral blood leukocytes (PBL) following stimulation with the eight Culicoides<\/jats:italic> allergens was measured in 28 IBH-affected and 24 healthy control horses. Allergen-specific serum IgE levels was determined by ELISA.<\/jats:p><\/jats:sec>Results<\/jats:title>The eight major allergens were successfully expressed in insect cells and purified. All allergens induced a significantly higher sLT release from PBL of IBH-affected horses compared to healthy controls. There was a high correlation and substantial to excellent agreement between sLT release and serum IgE levels for six Culicoides<\/jats:italic> allergens, while for two, the agreement was moderate. Positivity rates in IBH horses were usually higher in IgE serology, but more false-positive results were obtained. The allergens performing best in both assays were Cul o 3, Cul o 8, and Cul o 9, with very high specificity and good sensitivity.<\/jats:p><\/jats:sec>Discussion<\/jats:title>Insect-cell-expressed Culicoides<\/jats:italic> recombinant allergens are functionally relevant and will open new opportunities for the study of Culicoides<\/jats:italic> hypersensitivity not only in horses but also potentially in human patients or other species. They will also greatly improve IBH diagnostics using cellular assays and IgE serology.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1597233","type":"journal-article","created":{"date-parts":[[2025,6,6]],"date-time":"2025-06-06T04:10:44Z","timestamp":1749183044000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Culicoides allergens expressed in insect cells induce sulphidoleukotriene release from peripheral blood leukocytes of horses affected with insect bite hypersensitivity"],"prefix":"10.3389","volume":"16","author":[{"given":"Sigridur","family":"Jonsdottir","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara 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