Advertisement

Topics

Cloning and expression of candidate allergens from Culicoides obsoletus for diagnosis of insect bite hypersensitivity in horses.

08:00 EDT 15th June 2013 | BioPortfolio

Summary of "Cloning and expression of candidate allergens from Culicoides obsoletus for diagnosis of insect bite hypersensitivity in horses."

Insect bite hypersensitivity (IBH) is an IgE-mediated (Type I) hypersensitivity reaction induced by allergens from biting midges of the Culicoides spp. The aim of the present study was to identify, clone and express recombinant allergens from C. obsoletus, the main species found feeding on horses in the Netherlands, by sequence homology searches on the C. obsoletus specific RNA database, with previously described allergens from C. nubeculosus and C. sonorensis. BLAST searches with these described allergens resulted in similarity hits with 7 genes coding for C. obsoletus allergens. These allergens were expressed as hexahistidine tagged recombinant proteins in E. coli. Allergens were termed Cul o 1-Cul o 7. A maltase (Cul o 1) plus Cul s 1 (maltase of C. sonorensis) were additionally expressed in insect cells using the baculovirus expression system to compare homologous allergens from different species produced with different expression systems in diagnostic in vitro and in vivo tests. We demonstrate that IBH affected horses in the Netherlands show higher IgE levels to Cul o 1 than to Cul s 1, as determined by an IgE ELISA. Furthermore, we show that Cul o 1 produced in E. coli is at least as suitable for in vitro diagnosis of IBH affected horses as Cul o 1 produced in the baculovirus/insect cell expression system. The resulting proteins were evaluated for their ability to discriminate IBH affected and healthy horses by ELISA and intradermal testing. The frequency of positive test results by ELISA within IBH affected horses ranged from 38% to 67% for the different allergens. When results of IgE-binding to Cul o 1-Cul o 7 were combined the test had a sensitivity of 92% and specificity of 85%. The capability of the allergens to induce Type I hypersensitivity reaction in IBH affected horses was demonstrated by an intradermal test. The results show that E. coli expressed recombinant allergens from C. obsoletus are valuable tools to determine the allergen specific sensitisation profile (component resolved diagnosis) in horses with IBH in countries were C. obsoletus is the most abundant species and may facilitate in the development of future immunotherapy.

Affiliation

Cell Biology and Immunology Group, Wageningen University, Wageningen, The Netherlands.

Journal Details

This article was published in the following journal.

Name: Veterinary immunology and immunopathology
ISSN: 1873-2534
Pages: 227-39

Links

DeepDyve research library

PubMed Articles [19989 Associated PubMed Articles listed on BioPortfolio]

Three new Scandinavian species of Culicoides (Culicoides): Culicoides boyi sp. nov., Culicoides selandicus sp. nov. and Culicoides kalix sp. nov. (Diptera: Ceratopogonidae).

In the context of a major monitoring program of Culicoides in Denmark and Sweden due to the appearance of bluetongue disease in 2007-2008, a large number of specimens were collected by light traps and...

Seasonal Dynamics, Parity Rate, and Composition of Culicoides (Diptera: Ceratopogonidae) Occurring in the Vicinity of Wild and Domestic Ruminants in the Czech Republic.

In the light of the emergence of bluetongue and Schmallenberg viruses in northern and central Europe, an extensive entomological survey within the framework of a bluetongue control program was underta...

Dermatophagoides farinae allergen Der f 9: Cloning, expression, purification, characterization and IgE-binding in children with atopic asthma.

The house dust mite species Dermatophagoides farinae releases allergens that cause allergies and asthma worldwide. This study sought to clone and express the full-length cDNA encoding the group 9 alle...

Comparison of Vertebrate Cytochrome b and Prepronociceptin for Blood Meal Analyses in Culicoides.

To date, studies on host preferences and blood meal identification have been conducted for Culicoides species using molecular-based methods such as PCR techniques to amplify only a fragment from unive...

Evaluation of in vitro methods for assessment of infection of Australian Culicoides spp. with bluetongue viruses.

Biting midges from the genus Culicoides (Diptera: Ceratopogonidae) are the vectors of several globally important arboviruses that affect livestock. These include orbiviruses from the bluetongue virus ...

Clinical Trials [2925 Associated Clinical Trials listed on BioPortfolio]

Identification of Anaphylactogenic Antibodies in Peanut Allergy

This project intends the analysis and profiling of specific antibodies against major peanut allergens in peanut allergic individuals and molecular cloning of human antibodies against major...

Positional Cloning of the Gene(s) Responsible for Alagille Syndrome

The goal of the project is to identify and clone the gene(s) responsible for the Alagille Syndrome (AGS) by a positional cloning approach. The first step towards this goal is to define th...

Potential Allergens in Wine: Double-Blind Placebo-Controlled Trial

This study is designed to identify whether wines which are produced using the common potential food allergens such as proteins derived from fish, milk or egg are likely to contain sufficie...

Neutrophil FFA2/GPR43 Receptor Expression in Patients With the Diagnosis of Sepsis

This study seeks to elucidate the quantitative expression of G - protein receptor 43/free fatty acid (GPR43/FFA2) receptors in patients with the diagnosis of sepsis and specifically, its e...

Evaluating for Contact Allergies in Patients With Chronic Urticaria

Hives affects 10-25% of the population worldwide at some time during their lifetime. Hives are itchy transient swellings of the skin lasting 4-36 hours. Chronic urticaria is defined as hiv...

Medical and Biotech [MESH] Definitions

An in vitro allergen radioimmunoassay in which allergens are coupled to an immunosorbent. The coupled allergens bind the IgE in the sera of patients which in turn binds radioisotope-labeled anti-IMMUNOGLOBULIN E antibodies.

Methods pertaining to the generation of new individuals, including techniques used in selective BREEDING, cloning (CLONING, ORGANISM), and assisted reproduction (REPRODUCTIVE TECHNIQUES, ASSISTED).

Tests involving inhalation of allergens (nebulized or in dust form), nebulized pharmacologically active solutions (e.g., histamine, methacholine), or control solutions, followed by assessment of respiratory function. These tests are used in the diagnosis of asthma.

Application of allergens to the nasal mucosa. Interpretation includes observation of nasal symptoms, rhinoscopy, and rhinomanometry. Nasal provocation tests are used in the diagnosis of nasal hypersensitivity, including RHINITIS, ALLERGIC, SEASONAL.

Antigens from the house dust mites (DERMATOPHAGOIDES), mainly D. farinae and D. pteronyssinus. They are proteins, found in mite feces or mite extracts, that can cause ASTHMA and other allergic diseases such as perennial rhinitis (RHINITIS, ALLERGIC, PERENNIAL) and atopic dermatitis (DERMATITIS, ATOPIC). More than 11 groups of Dermatophagoides ALLERGENS have been defined. Group I allergens, such as Der f I and Der p I from the above two species, are among the strongest mite immunogens in humans.

Quick Search
Advertisement
 


DeepDyve research library

Relevant Topic

Cloning
Cloning in biology is the process of producing similar populations of genetically identical individuals that occurs in nature when organisms such as bacteria, insects or plants reproduce asexually. Cloning in biotechnology refers to processes used to cre...


Searches Linking to this Article