Chinese expert consensus on allergen component resolved diagnosis
Wenting LuoHao ChenLei ChengYubao CuiYinshi GuoZhongshan GaoKai GuanKun HanHaiyu HongKunmei JiJing LiGuanghui LiuJuan MengJinlyu SunAilin TaoWei TangHuiying WangXiaoyan WangJi‐Fu WeiXuejun ShaoLi XiangStephen Kwok‐Wing TsuiHuanping ZhangYongmei YuLan ZhaoZhifeng HuangHui GanJiale ZhangXianhui ZhengPeiyan ZhengHuimin HuangChuangli HaoRongfei ZhuBaoqing Sun
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Abstract:
Allergen component resolved diagnosis (CRD) is a method for identifying specific protein molecules that cause hypersensitivity. Unlike traditional methods that use crude allergen extracts containing multiple component species, CRD focuses on individual allergen protein molecules for more precise diagnosis. The World Allergy Organization (WAO) recommends CRD as a supplement to clinical history and allergen extract testing, and in some cases, it can replace crude extract tests. CRD involves the use of natural or recombinant proteins to detect specific IgE antibodies directed at individual allergenic components. This method allows for a more detailed analysis of a patient's allergic response compared to the use of whole allergen extracts. The Allergy Prevention and Control Specialty Committee of the Chinese Preventive Medicine Association, in collaboration with multidisciplinary experts, developed an expert consensus that incorporates the consensus of the European Academy of Allergy and Clinical Immunology (EAACI), WAO, and important domestic literature on CRD in recent years. The consensus aims to standardize the algorithm of allergen diagnosis and provides a reference for clinical practice. It also offers guidance for clinicians on the common protein families identified by CRD, the scenarios where CRD is applicable, and the significance of detecting common allergen components. Despite its potential, CRD is not widely used in clinical practice in China due to the lack of allergen component reagents and a general unawareness among clinicians about CRD's application and interpretation of test results. The expert consensus developed by the Chinese Preventive Medicine Association aims to address this gap and enhance the clinical application of CRD in China.Keywords:
Clinical Practice
【Objective】 To understand the proportion of allergen in eczema of children for providing foundation for prevention and treatment of eczema.【Method】 Bioresonance technology allergy therapy was used to detect the allergen of eczema in children.【Results】 A total of 1 538children with eczema were detected,1 272(82.70%)patients,among them had positive food allergen test.653(42.46%)patients had positive inhaled allergen test.504(32.77%)patients had positive allergen test both in food test and inhaled test.Cereal(1 020,66.32%)was the most common allergen in food allergen,followed by milk,egg,sea food,nut,bean,food preservative,beef and lamb;Dust(340,22.11%)was the most common allergen in inhaled allergen,followed by mite,flower,animal fur,mould,waste gas,cigarettes,cooking oil fume.Among different aging groups,there were significant differences in the positive rates between food allergen group and inhaled allergen group(P0.05).Food allergen was the main allergen in infant eczema,with the growth of the age,inhaled allergen was the main allergen in elder children eczema.【Conclusion】 The inducing factor of eczema can be found out by the detection of allergen,which providing the foundation for the prevention and treatment of eczema.
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Food allergens
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Natural allergen contact induces an increase of IgE levels and sensitivity but the mechanisms underlying the allergen-specific memory responses are poorly understood. Furthermore, it has not been studied whether allergen exposure affects the molecular reactivity profiles in patients. The aim of this study was to analyze the influence of nasal allergen encounter on the molecular profile and magnitude of memory IgE responses and on systemic sensitivity.We investigated allergen-specific IgE, IgG subclass and IgM responses to defined allergen molecules (grass pollen: Phl p 1, Phl p 2 and Phl p 5; birch pollen: Bet v 1 and Bet v 2) in allergic patients in response to natural as well as to controlled nasal and dermal allergen exposure. Changes in systemic sensitivity were monitored by skin prick testing and by basophil histamine release experiments.Respiratory antigen exposure boosted IgE levels to a pre-established profile of allergen molecules without inducing significant IgM responses or new IgE specificities in allergic individuals. The importance of the route of allergen contact is demonstrated by an increase of systemic IgE levels and sensitivity after nasal exposure. In vitro sensitisation of basophils with pre- and post-seasonal serum samples suggests an allergen-induced elevation of specific IgE as a cause for the increased allergen-specific sensitivity.The characteristics of the allergen-driven antibody responses indicate a direct activation of an established pool of IgE memory cells with defined specificities as an underlying mechanism. Our finding that nasal allergen contact is a major factor for the boosting of memory IgE and systemic sensitivity may open new therapeutic possibilities.
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Commercial extracts used for diagnosis and treatment of allergy are currently prepared from natural allergen sources. The aim of this study was to analyse birch pollen allergen extracts produced for in vivo diagnosis of birch pollen allergy regarding their contents of individual birch pollen allergens (Bet v 1, Bet v 2 and Bet v 4).Protein contents were measured and the allergen composition was analysed by immunoblotting using antibody probes specific for Bet v 1, Bet v 2 and Bet v 4 in birch pollen extracts from five manufacturers of allergen extracts. The contents of the major birch pollen allergen, Bet v 1, were quantified with a specific two-site binding enzyme-linked immunosorbent assay with nanogram sensitivity for Bet v 1. The biological activities of the allergen extracts were evaluated by skin prick testing in birch pollen allergic patients and compared with their sensitization profiles.A more than 10-fold variation regarding total protein contents (23.1-314 microg mL(-1)) and also regarding the amounts of the major birch pollen allergen, Bet v 1 (1.62-19.6 microg mL(-1)) was found. The highly cross-reactive Bet v 4 allergen was absent in three of the five tested extracts. Furthermore, varying skin test results were obtained in birch pollen allergic patients with the allergen extracts.Commercial birch pollen extracts exhibit a considerable variability regarding allergen contents and hence deliver varying in vivo test results. These problems might be overcome with recombinant allergen-based preparations.
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Seven monoclonal antibodies (mAbs) against the wheat allergen, Tri a Bd 17 K, were prepared to obtain mAbs suitable for a sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for determination of the allergen. Two of the mAbs strongly immunoblotted the allergen purified from wheat flour. However, only one (1G11) of them was found to be suitable for sandwich ELISA. Epitope mapping against mAb-1G11 on the allergen showed that the mAb recognized the peptide containing Lys-38 and Gln-39 of the allergen. We developed a sandwich ELISA method consisting of Aleuria aurantia lectin for fixing the allergen and 1G11 as the first antibody that enabled 4-4,000 ng/well of the allergen to be determined.
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Allergen extracts and recombinant allergens are used in allergy diagnostics and immunotherapy. Since allergen extracts from different manufacturers lack proper standardization regarding their composition, monoclonal antibodies (MAbs) against specific allergen components can be used for their identification and quantification in allergen extracts. This study aimed to generate MAbs against allergen Der p 21 of Dermatophagoides pteronyssinus for the analysis of allergen extracts. Recombinant Der p 21 was expressed in E. coli and purified using affinity chromatography. MAbs against Der p 21 were generated using hybridoma technology. House dust mite (HDM) allergen extracts were analyzed using the newly developed sandwich enzyme-linked immunosorbent assay, Western blotting and microarray immunoassay. MAbs were proven to be reactive with natural Der p 21. Highly specific sandwich enzyme-linked immunosorbent assay for the quantification of Der p 21 was developed. The allergen was detected and its concentration was determined in only 3 of 6 analyzed HDM allergen extracts from different manufacturers. HDM analysis by MAb-based immunoassays shows their differences in allergen composition. The results demonstrate the importance of allergen-specific MAbs as a tool for the characterization of allergen extracts and the need for their appropriate standardization before their use for allergy diagnostics or immunotherapy.
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The major allergen (Allergen M) from codfish is an acidic protein, mol. wt. 15000 with 114 amino acid residues and only one sugar (glucose) residue. It is highly active in skin testing on codfish-allergic individuals, in passive transfer (PK) testing and in the radioallergosorbent test (RAST). The activity resists denaturation and incomplete tryptic hydrolysis. Two fragments (TM 1, mol. wt. 8500, and TM 2, mol. wt. 6500) are obtained from allergen M. The fragments are equally active and allergenically identical in biological tests, but somewhat less active than Allergen M itself. Smaller peptides obtained from TM 2 are inactive. The amino acid sequence of TM 2 is given as the first reported example of a full structural analysis of an active allergen. The information obtained in studies of this model allergen is compared with information published from other studies of allergens. Similarities and differences are pointed out.
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