期刊
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
卷 137, 期 1-2, 页码 76-83出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.vetimm.2010.04.012
关键词
Horse; Insect bite hypersensitivity; Simulium vittatum; Gulicuides nubeculosus; Sim v 1; Cul n 1; Cross-reactivity
资金
- Swiss National Science Foundation [320030_127618/1, 31-116803/1]
- Department of Clinical Veterinary Medicine of the Vetsuisse Faculty of Berne
- Agricultural Productivity Fund of Iceland
- Swiss National Science Foundation (SNF) [320030_127618] Funding Source: Swiss National Science Foundation (SNF)
Insect bite hypersensitivity (IBH) is an IgE-mediated seasonal dermatitis of the horses associated with bites of Simulium (black fly) and Culicoides (midge) species. Although cross-reactivity between Simulium and Culicoides salivary gland extracts has been demonstrated, the molecular nature of the allergens responsible for the observed cross-reactivity remains to be elucidated In this report we demonstrate for the first time in veterinary medicine that a homologous allergen, present in the salivary glands of both insects, shows extended IgE cross-reactivity in vitro and in vivo The cDNA sequences coding for both antigen 5 like allergens termed Sim v 1 and Cut n 1 were amplified by PCR, subcloned in high level expression vectors, and produced as [His](6)-tagged proteins in Eschenchia coh. The highly pure recombinant proteins were used to investigate the prevalence of sensitization in IBH-affected horses by ELISA and their cross-reactive nature by Western blot analyses, inhibition ELISA and intradermal skin tests (IDT) The prevalence of sensitization to Sim v 1 and Cul n 1 among 48 IBH-affected horses was 37% and 35%, respectively In contrast, serum IgE levels to both allergens in 24 unaffected horses did not show any value above background. Both proteins strongly bound serum IgE from IBH-affected horses in Western blot analyses, demonstrating the allergenic nature of the recombinant proteins Extended inhibition ELISA experiments clearly showed that Sim v 1 in fluid phase is able to strongly inhibit binding of serum IgE to solid phase coated Cut n 1 in a concentration dependent manner and vice versa. This crucial experiment shows that the allergens share common IgE-binding epitopes. IDT with Sim v 1 and Cul n 1 showed clear immediate and late phase reactions to the allergen challenges IBH-affected horses, whereas unaffected control horses do not develop relevant immediate hypersensitivity reactions. In some horses, however, mild late phase reactions were observed 4h post-challenge, a phenomenon reported to occur also in challenge experiments with Simulium and Cuhcoides crude extracts probably related to lipopolysaccaride contaminations which are also present in E colt-expressed recombinant proteins. In conclusion our data demonstrate that IgE-mediated cross-reactivity to homologous allergens, a well-known clinically relevant phenomenon in human allergy, also occurs in veterinary allergy (C) 2010 Elsevier B V All rights reserved.
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