The Diagnostic Importance of Recombinant Allergen IgE Testing in Patients with Hymenoptera Venom Allergy: Comparison of Two Methods

Adults with systemic anaphylactic reactions (SAR) to insect sting show often multiple-positivity of serum-specific IgE (sIgE) to Hymenoptera venoms. Unnecessary long-lasting venom-specific immunotherapies (VIT) in false-positive patients increase the risk of recurrent SAR. This report aims to analyze the diagnostic importance of recombinant allergen IgE testing in patients with SAR to Hymenoptera sting.In 82 patients we measured sIgE to honeybee venom (HBV), wasp venom (WV) and hornet venom (HV) extracts, recombinant phospholipase A2 from HBV (sIgE-rApi m1), recombinant antigen 5 from WV (sIgE-rVes v5), and cross-reactive carbohydrate determinants-CCD-bromelain by ImmunoCAP. We analyzed the correlation of ImmunoCAP and Immunoblot for HBV and WV extracts, rApi m1, and rVes v5 in 39/82 patients. According to the history of the culprit insect, we compared sensitivity and specificity between the two methods.The severity of the SAR does not depend on the sIgE level to venom extracts and recombinant allergens. Fifty-one percent of the patients had a multiple-positivity to HBV/WV or HBV/WV/HV extracts. Severe SAR and CCD-sIgE were more frequent in multiple-positive than single-positive patients. CCD-sIgE were more frequent in HBV allergic patients than WV and HV allergic patients. There was a significant correlation between levels of sIgE to venom extracts and recombinant allergens measured by ImmunoCAP and Immunoblot. ImmunoCAP has higher sensitivity and specificity than Immunoblot for diagnosis of SAR to Hymenoptera venoms.IgE testing to recombinant CCD-free allergens is necessary for the adequate selection of long-lasting VIT, especially in patients with multiple sensitivities to venom extracts.

The importance of component-resolved diagnostics in IgE-mediated cow's milk allergy

Cow’s milk allergy (CMA) is an increasingly common problem among children and adults that requires the use of appropriate diagnostics to eliminate allergic reactions and prevent unnec-essary dietary regimes. The current diagnostics methods are imperfect hence new, more effective methods are still being sought. Component-resolved diagnostics (CRD) is one of them. CRD assesses sensitivity to individual allergen molecules using purified native or recombinant allergens. The present paper reviews the role of CRD in diagnosing CMA, as well as the benefits and limitations of its use, especially in predicting allergy development or acquiring immunotolerance. It examines the possibility of replacing the current gold diagnostic standard with component tests directed against specific milk proteins. In addition, CRD could be helpful in the evaluation of prognosis. However, CRD allows for improvement in clinical management, particularly of polysensitized subjects, there is still no cogent evidence that it offers more efficient CMA diagnostics than existing tests.

DNA vaccines and recombinant allergens with reduced allergenic activity treat allergies

This review is intended to familiarize readers with major novel directions of developing allergy vaccines, their structure, as well as the mechanisms of forming a new immunological response in the course of the treating immunoglobulin E (IgE)-mediated allergic diseases. Currently, science offers a huge variety of new experimental forms of recombinant allergens with reduced allergenic activity and increased immunogenicity, or vice-versa, immune tolerance. Often, the mechanisms of their effect on the immune system are not fully understood. Scientific publications, including reviews covering this topic, allowed us identifying top priority areas in the development of allergy vaccines: recombinant hypoallergenic allergen derivatives, T cell epitope-based allergy vaccines, and B cell epitope-based allergy vaccines. In addition, the review discusses use of deoxyribonucleic acid (DNA) vaccines. Immunotherapy with DNA vaccines is the newest and least studied method of treating allergic diseases.

Identification the Cross-Reactive or Species-Specific Allergens of Tyrophagus putrescentiae and Development Molecular Diagnostic Kits for Allergic Diseases

Mite allergens are considerable factors in the genesis of allergic diseases. The storage mite Tyrophagus putrescentiae (Tp) appears in contaminated foods and household surroundings. The current diagnostic tools for Tp allergy are mostly based on crude extracts and still contain shortcomings. This study aimed to investigate the immunoglobulin E (IgE)- responsiveness profiles of Tp-allergic patients and develop a molecular diagnostic method using recombinant allergens. Allergenic components were characterized as cross-reacting or species-specific allergens, in which the effective combinations of recombinant allergens were developed and analyzed in terms of the prediction accuracy for clinical diagnosis. Seven recombinant allergens were cloned and generated to detect the IgE responsiveness of the Tp allergy. A survey on the prevalence of mite allergy showed there were higher sensitizations with IgE responsiveness to house dust mites (HDM) (78.9–80.9%) than to storage mites Tp (35.6%). Prevalence of sensitization to Tp was higher in elderly subjects. The principal IgE-binding components of Tp were Tyr p 1, Tyr p 2 and Tyr p 3. Prediction accuracy for Tp allergy by IgE-responsiveness combination D (Tyr p 1, Tyr p 2 & Tyr p 3) was with high precision (100%). Avoiding the cross-reactivity of Dermatophagoides pteronyssinus, the prediction accuracy of IgE-responsiveness combination H+ (Tyr p 1, Tyr p 2, Tyr p 3, Tyr p 7, Tyr p 8, Tyr p 10 & Tyr p 20) was suitable for Tp-specific diagnosis. Panels of Tp allergens were generated and developed a diagnostic kit able beneficial to identify IgE-mediated Tp hypersensitivity.

Recombinant Allergens in Immunotherapy of Asthma

Asthma is a chronic inflammatory disorder caused by T-cell-mediated inflammation within airways. The prevalence of allergic diseases is rapidly increasing so that knowing allergens (characterization and types) and strategies for asthma management, prevention and treatment are very important. The most strategy is the production of recombinant allergens. Many of the problems associated with using natural allergenic products for allergy diagnosis and treatment can be overcome with the use of genetically engineered recombinant allergens. Various recombinant allergens are now emerging as strong candidates for allergen-specific immunotherapy. Extrinsic asthma (a type of asthma) may respond to immunotherapy such as using recombinant allergens. These exciting novel therapies provide not only the promise of new therapies for asthma but also valuable tools for the investigation of asthma mechanisms. This review describes strategies for asthma management, prevention and treatment, and especially recombinant allergens and also recent progresses in the molecular biology of recombinant allergens and then advantage and disadvantage of these allergens are explained. There are many methods for producing allergens such as extraction of serum, Ro/SS-A anti-Ro/SS-A system, Using the Solid-Phase Immunoadsorption system and finally recombinant technology for producing recombinant allergens. Recombinant allergens can express in many systems such as bacteria, yeast, insect cells, animal cells, and transgenic plant. We describe recombinant allergens produced in these systems. The obtained results hold promise that recombinant allergen–based immunotherapy will improve current immunotherapy practice and may open possibilities for new treatment strategies and possibly even for prophylactic vaccination.