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Cloning, production and characterization of antigen 5 like proteins from Simulium vittatum and Culicoides nubeculosus, the first cross-reactive allergen associated with equine insect bite hypersensitivity.
Vet Immunol Immunopathol. 2010 Sep 15; 137(1-2):76-83.VI

Abstract

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 Cul n 1 were amplified by PCR, subcloned in high level expression vectors, and produced as [His](6)-tagged proteins in Escherichia coli. 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 Cul 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 Culicoides crude extracts probably related to lipopolysaccaride contaminations which are also present in E. coli-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.

Authors+Show Affiliations

Swiss Institute of Allergy and Asthma Research (SIAF), University of Zürich, Obere Strasse 22, CH-7270 Davos, Switzerland.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20537727

Citation

Schaffartzik, A, et al. "Cloning, Production and Characterization of Antigen 5 Like Proteins From Simulium Vittatum and Culicoides Nubeculosus, the First Cross-reactive Allergen Associated With Equine Insect Bite Hypersensitivity." Veterinary Immunology and Immunopathology, vol. 137, no. 1-2, 2010, pp. 76-83.
Schaffartzik A, Marti E, Crameri R, et al. Cloning, production and characterization of antigen 5 like proteins from Simulium vittatum and Culicoides nubeculosus, the first cross-reactive allergen associated with equine insect bite hypersensitivity. Vet Immunol Immunopathol. 2010;137(1-2):76-83.
Schaffartzik, A., Marti, E., Crameri, R., & Rhyner, C. (2010). Cloning, production and characterization of antigen 5 like proteins from Simulium vittatum and Culicoides nubeculosus, the first cross-reactive allergen associated with equine insect bite hypersensitivity. Veterinary Immunology and Immunopathology, 137(1-2), 76-83. https://doi.org/10.1016/j.vetimm.2010.04.012
Schaffartzik A, et al. Cloning, Production and Characterization of Antigen 5 Like Proteins From Simulium Vittatum and Culicoides Nubeculosus, the First Cross-reactive Allergen Associated With Equine Insect Bite Hypersensitivity. Vet Immunol Immunopathol. 2010 Sep 15;137(1-2):76-83. PubMed PMID: 20537727.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning, production and characterization of antigen 5 like proteins from Simulium vittatum and Culicoides nubeculosus, the first cross-reactive allergen associated with equine insect bite hypersensitivity. AU - Schaffartzik,A, AU - Marti,E, AU - Crameri,R, AU - Rhyner,C, Y1 - 2010/04/28/ PY - 2010/01/22/received PY - 2010/04/08/revised PY - 2010/04/19/accepted PY - 2010/6/12/entrez PY - 2010/6/12/pubmed PY - 2010/12/14/medline SP - 76 EP - 83 JF - Veterinary immunology and immunopathology JO - Vet Immunol Immunopathol VL - 137 IS - 1-2 N2 - 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 Cul n 1 were amplified by PCR, subcloned in high level expression vectors, and produced as [His](6)-tagged proteins in Escherichia coli. 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 Cul 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 Culicoides crude extracts probably related to lipopolysaccaride contaminations which are also present in E. coli-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. SN - 1873-2534 UR - https://www.unboundmedicine.com/medline/citation/20537727/Cloning_production_and_characterization_of_antigen_5_like_proteins_from_Simulium_vittatum_and_Culicoides_nubeculosus_the_first_cross_reactive_allergen_associated_with_equine_insect_bite_hypersensitivity_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0165-2427(10)00127-3 DB - PRIME DP - Unbound Medicine ER -