Recent advances in the study of epitopes,allergens and immunologic cross-reactivity of edible mango

Mango(Mangifera indica L.)is a tropical fruit that is widely consumed as both fresh fruits and processed products around the world.The high incidence of mango allergy,on the other hand,has sparked widespread concern.Therefore,a summary and analysis of the current status and issues in mango allergen research can guide in-depth study on the mechanism of mango allergy and reveal effective desensitization methods.We described the incidence of fruit allergy,as well as the mechanism and clinical symptoms of mango allergy,in this review.We also looked into the structural properties of mango allergens,the effect of processing methods on mango allergens,prediction methods for mango allergen epitopes,and the current state of research on mango cross-reactive allergens and preventive measures.Finally,the research directions and ideas for the future are proposed and discussed.

Validation of Molecular Detection Methods for Gluten Allergens in Infant Formula

[Objectives]To verify the specificity,sensitivity,precision and negative-positive deviation of the foodproof gluten component de-tection kit for the detection of gluten allergens in milk powder matrix,and to establish a real-time fluorescent PCR legal method for the detec-tion of gluten allergens in milk powder.[Methods]The specificity,sensitivity,precision and negative-positive deviation of the detection method of foodproof gluten component detection kit(PCR-probe method)were verified by artificially adding different concentrations of wheat bran and extracting sample DNA by kit method,and applied to sample detection.[Results] The specific detection results of two kinds of milk powder with wheat bran and buckwheat added showed that the foodproof gluten component detection kit(PCR-probe method)had good speci-ficity for wheat gluten.The results of artificially added wheat bran positive samples showed that the false positive rate and false negative rate of the kit in the milk powder matrix were O,and the sensitivity and precision were high.[Conclusions]The kit is simple to operate and has high accuracy,which is suitable for the detection of gluten allergen components in milk powder.

The detection and regulation advances of food allergens

Food allergy has become an important food quality and safety issue,posing a challenge to the food industry and affecting consumer health.On the one hand,from the perspective of food processing industry,the diversity of food raw material ingredients,exogenous additives,and processing forms make the presence of allergens in modern food processing more complex.In addition,due to the lack of allergen identification,effective detection and allergenicity evaluation systems,there are serious deficiencies in the current theories and techniques for food allergen screening and detection,tracking and prediction,intervention and control;On the other hand,from the perspective of public health,meeting consumers'right to know whether there are raw materials containing food allergens in processed foods,and improving the credibility of government and people's satisfaction have become urgent matters;In addition,as people come into contact with more and more new borne novel foods,the probability of food allergy is also increasing.The food safety and health problems induced by increasingly complex,widespread and severe food allergy are difficult to avoid.In view of this,in response to the increasingly serious food allergy issues,this paper introduced the detection methods of food allergens,summarized the reduction and control techniques of food allergens,and elaborated hypoallergenic foods,which aims to provide the basis for preventing and controlling food allergy and ensuring the physical health of food allergy patients.

Allergenicity mitigation strategies of seafood allergens for hypoallergen

Introduction Seafood is a widely consumed and nutritionally rich food source,but it ranks among the top allergens,with shellfish and fish being major triggers of allergic reactions.These reactions,primarily caused by proteins such as tropomyosin(TM)in shellfish and parvalbumin(PV)in fish,can lead to life-threatening anaphylaxis in susceptible individuals.The development of hypoallergenic seafood is becoming increasingly important to provide safe alternatives for those with seafood allergies.This comment reviews recent advances in allergenicity mitigation strategies,highlighting the progress made and future potential in producing hypoallergenic seafood products.

Analysis of Allergens in 5473 Patients with Allergic Diseases in Harbin, China

Objective To analyze the allergic status to common inhalant allergens and food allergens in clinical patients in Harbin in northeastern China and provide evidence to develop the prevention strategy of allergic disease. Methods The data were collected from 5 473 patients with clinical suspected allergic diseases seeking medical care in the Second Affiliated Hospital of Harbin Medical University. Among these patients, 2 530 （46.2%） were males aged 0-86 years, the youngest was only 1 month old and 2 579 （47.1%） were young children and teenagers. The serum specific Immunoglobulin E （slgE） to 14 kinds of common allergens and serum total IgE were detected by using AllergyScreen test （Mediwiss Analytic GmbH, Moers, Germany）. Results In 5 473 subjects the positive rate of slgE was 33.1% （n=1 813）. Cow milk （6.9%） and wheat （3.1%） were the most common food allergens, followed by house dust mite mix （12.5%） and mould mix （9.4%） and the age and gender specific differences in the positive rate were significant. For the children aged 〈7 years the positive rates to cow milk, beef-mutton, and egg white/egg yolk were high, but the positive rates to house dust mite mix, ragweed estragon, and mould mix were low （P〈0.05）. For the adults the positive rates to aeroallergens were high while the rates to food allergens were low. Conclusion The results from this study showed that the food allergens in Harbin had geographic characteristics, which support the viewpoint that the environment factors play an important role in the incidence of allergic diseases. Also, the detection of slgE and total IgE are essential to identify relevant allergens for the purpose of early diagnosis, management and prevention of allergic disease.

A Study on Pollen Allergens in China

Objective Allergic disease caused by airborne pollen is a major health problem in China. Intensive study on pollen allergens can be of great help for preventing and treating pollinosis. Four aspects of the study on pollen allergens in China including major allergic pollen in our country, analysis and purification of pollen allergen composition, recombinant pollen allergens and clinical application of pollen allergens are described in this paper.

Cross-reactivity between aeroallergens and food allergens

In patients with respiratory allergy,cross-reactivity between aeroallergens and foods may induce food allergy,symptoms ranging from oral allergy syndrome to severe anaphylaxis.Clinical entities due to Ig E sensitization to cross-reactive aeroallergen and food allergen components are described for many sources of plant origin(pollen-food syndromes and associations,such as birch-apple,cypress-peach and celery-mugwortspice syndromes,and mugwort-peach,mugwortchamomile,mugwort-mustard,ragweed-melon-banana,goosefoot-melon associations),fungal origin(Alternariaspinach syndrome),and invertebrate,mammalian or avian origin(mite-shrimp,cat-pork,and bird-egg syndromes).Clinical cases of allergic reactions to ingestion of food products containing pollen grains of specific plants,in patients with respiratory allergy to Asteraceae pollen,especially mugwort and ragweed,are also mentioned,for honey,royal jelly and bee polen dietary supplements,along with allergic reactions to foods contaminated with mites or fungi in patients with respiratory allergy to these aeroallergens.Medical history and diagnosis approach may be guided by the knowledge about the diverse cross-reacting allergens involved,and by the understanding of these clinical entities which may vary significantly or may be overlapping.The association between primary Ig E sensitization with respiratory symptoms to inhaled allergens and food allergy due to cross-reactive allergen components is important to assess in allergy practice.The use of molecular-based diagnosis improves the understanding of clinically relevant Ig E sensitization to cross-reactive allergen components from aeroallergen sources and foods.

Detection and inactivation of allergens in soybeans:A brief review of recent research advances

In the last decade,soybean allergies have been on the increases to such an extent that they have now become a public health issue thus prompting more studies and researches on the topic.The allergenicity of soybean is attributed to its protein fraction.The best way to prevent hypersensitive patients from ingesting allergenic compounds is to exclude such soybean allergens from their diet.As a result,it is essential to provide detailed and reliable knowledge of food ingredients.Therefore,precise and reliable approaches for detecting soybean allergens found in various food products must be used.The main way to reduce allergy risk is the identification of allergenic sites in food and their inactivation by various food-processing methods.It has been reported that food processing may lead to the modification of conformational structure of the protein or protein distortion that inhibit the binding of immunoglobulin E(Ig E)to epitopes on food allergens and also the mechanism of allergic reactions.Food processing technologies employed for inactivating allergenic epitopes used thermal and nonthermal techniques.Currently,several detection methods including protein-based and DNA-based approaches using analytical techniques such as enzyme-linked immunosorbent assay(ELISA),enzyme allergosorbent test(EAST),radioallergosorbent test(RAST),lateral flow immunoassay(LFIA),immunoblotting,realtime polymerase chain reaction(PCR),mass spectrometry and biosensors have been improved for identifying and quantifying these epitopes.This research focused on allergenic proteins of soybean,the most modern approaches for detecting and quantifying these allergens,and finally,the various methods used to inactivate these proteins and their effects on soy allergenicity.

IgE Reactivity of Potential Allergens from the Swimming Crab Portunus trituberculatus and the Research of ELISA Reagent for Detecting Crab Food Anaphylaxis

Crustacean is one of the major allergic foods.It is of great significance to identify more crab allergens and research the detection methods for crab food anaphylaxis.In this study,IgE reactivity to three recombinant proteins from Portunus trituberculatus,including tropomyosin(rPtTM),myosin light chain(rPtMLC),and pancreatic lipase(rPtPL),were detected by enzyme-linked im-munosorbent assay(ELISA).The expressions of tropomyosin(TM)in various tissues of P.trituberculatus were detected by Western blot(WB).Furthermore,microplates were coated with rPtTM and the ELISA conditions were optimized.The cut-off value was deter-mined by the receiver operating characteristic(ROC)curve.Among 51 crab-allergic sera,21(41.2%)showed positive IgE to rPtTM.Other 70 crab-allergic sera more frequently recognized rPtPL(9/70;12.9%),followed by rPtMLC(1/70;1.4%).WB results showed that TM was mainly expressed in the muscle,followed by the heart and a small amount in gills and the testis.The optimal results showed that the coating condition of rPtTM was 50 ng per well with coating for 3 h at 37℃.The optimal blocking condition was 1.2%BSA with blocking for 3 h at 37℃,and the optimal dilution of the second antibody was 1:1500.The ROC curve showed that the ELISA reagent had high sensitivity(83.52%)and specificity(98.00%)when the cut-off value was 0.45.All results indicated that tropomyosin is the major allergen of P.trituberculatus,and myosin light chain and pancreatic lipase are the potential allergens.Addi-tionally,the ELISA reagent developed with the rPtTM was feasible for laboratory detection of crab anaphylaxis.

Food Allergens and Food Safety： A Global Perspective with Respect to Codex Alimentarius

Over the last two decades, food allergens are being recognized as one of the food hazards. This serious food safety issue is being addressed in different countries not only in terms of their biological and clinical characteristics, but also in terms of various standards of allergen management in food industries. This abnormal immune response caused by food allergens affects the quality of life especially in children and influences their overall health and retards normal growth, along with they suffer from ailments like eating disorders, depression, sometimes even death which is the most adverse impact of food allergy. Every country has their own set of guidelines to deal with the food allergens especially developed countries, food allergen guidelines is not well documented in developing countries. FAO/WHO is providing assistance to developing countries in strengthening their food safety guidelines. The Codex Alimentarius was formulated by the two organizations FAO/WHO in the year 1960; it is a collection of food standards in a integrated, codified style, together with associated material such as codes of hygiene and good manufacturing practices that should be followed by industries during various production stages of a food item. In both developed and developing nations, the Food Chemical Codex or the Codex Alimentarius aims to protect public health and implement fair trade practises for the trading of food items.

Induced Immunity of Response to Some Allergens Towards Response of Other Skin Test Allergens and Its Correlation with Immediate Hypersensitivity Reactions

Allergens are innocuous environmental antigens that cause lgE mediated hypersensitivity reactions. These antigens can be divided into several groups according to their nature like plant allergens, grasses, fungal allergens, pollen grains and house dust mite. Skin response to such allergens is associated with each other either by positive relation （positive correlation） or negative （inverse） relation. No previous papers found to discuss such correlations. In order to investigate these correlations, skin prick test of standard allergens solutions was carried out in 500 patients suffering allergic reactions of the age 12-60 years of both sexes. The results showed that there is a high significance of positive and negative correlations in skin response to allergens in the same group and with allergens of different groups suggesting that there is an induction or suppression of immune response. In conclusion the similarity or dissimilarity in the primary structure of some allergens in addition to gene-environment interaction may reflect positive/negative correlations between certain allergens that can influence the related allergic reactions.

Peanut Allergens Attached with p-Aminobenzamidine Are More Resistant to Digestion than Native Allergens

Despite being known as resistant proteins, peanut allergens (Ara h 1 and Ara h 2) can be digested and cause allergic reactions. Making the allergens more resistant to digestion may aid in non-absorption and excretion of the allergens. Our objectives were to make Ara h 1 and Ara h 2 more resistant to digestion and test them in a model system using trypsin as the digestive enzyme. The resistant allergens were prepared by covalently attaching p-aminobenzamidine (pABA), a protease inhibitor, to peanut allergens in an extract or on a PVDF membrane using glutaraldehyde, and were then tested for resistance to trypsin digestion. SDS-PAGE and Western blot were performed to determine the allergenic capacity of the modified allergens. A control was prepared using glycine instead. Results showed that Ara h 2, when covalently attached with pABA, was more resistant to trypin digestion than the native allergen. Similarly, Ara h 1, prepared on a PVDF membrane and treated with pABA, displayed a resistance to trypsin digestion. Treatment of the allergens with glycine (a control) instead of pABA showed that the modified allergens were as digestible as native allergens. Blot assays showed that the pABA-treated allergens exhibited a lower allergenic capacity than native allergens. It was concluded that pABA, when attached to peanut allergen Ara h 1 or Ara h 2, inhibited digestion of the allergen by trypsin and reduced their allergenic capacity as well.

In Silico Analysis of the Cross-Reactivity of Pacific White Shrimp (Litopenaeus vannamei) Allergens with Other Arthropod Species

This study provided new data in the area of cross-reactivity for Pacific white shrimp (Litopenaeus vannamei). Although the cross-reactivity between the most prominent allergenic proteins of shrimp and other crustaceans has been extensively studied, few data are showing the frequency of arthropod-shrimp Litopenaeus vannamei cosensitization in an in silico analysis. A comparative analysis of “typical and non-typical” shrimp L. vannamei allergens with other species was achieved, revealing new allergens and previously characterized shrimp allergens, which validated the comprehensive identification approach used in this study. Importantly, up to 192, amino acid sequences were identified that had matches to shrimp L. vannamei allergens that matched allergenic proteins in mites, insects, fish, bacteria, mammals, birds, and plants.

Determination of 24 Allergens in Perfume with High Performance Liquid Chromatography Tandem Mass Spectrometry

A method of 24 allergens determination in cosmetics were established with high performance liquid chromatography tandem mass spectrometry. The targeted compounds were extracted with acetonitrile and determined with LC-MS/MS (MRM mode) with external method. The linearity between concentrations and peak area ratio was obtained from 1.0~5.0 mg/L. The limits of detection were 1.0 mg/L for the instrument and 5.0 mg/kg for the method respectively. The LOQ was 15.0 mg/L. The average recoveries of 24 allergens were between 85.9% and 110.0% at spiked levels of 5, 10 and 20 mg/kg with relative standard derivation (RSDs) of 5.5%~12.0%(n=10). The method could be used as a reliable means for simultaneous quantitative determination of allergens in cosmetics.

Major allergens in 8 types of allergenic foods and allergen detection techniques

In recent years,the prevalence of allergens in food warning notices,both domestically and internationally,has become the second leading concern after microbial contamination.Among the various factors that threaten human health reported by the World Health Organization,food allergy ranks fourth,and food allergy has become a global security problem.As of now,no definitive treatment for food allergies exists,making the avoidance of allergen-containing foods the most effective prevention method.Consequently,labeling foods with allergen information serves as a crucial warning for allergic populations.Moreover,to enhance comprehension of food allergies,this article provides a brief overview of their definition and sensitization mechanisms.The main focus lies in highlighting the structure of primary allergens found in eight commonly allergenic foods and the resulting clinical symptoms they cause.Additionally,a summary of commonly employed allergen detection techniques is presented,with an analysis of their principles,advantages,and limitations.Looking ahead,the integration of diverse technological approaches to establish an efficient,accurate,and affordable allergen detection method remains a significant trend.This article has certain reference value for understanding the direction of food allergies.

Characterization of physicochemical and immunogenic properties of allergenic proteins altered by food processing:a review

Food allergens are mainly naturally-occurring proteins with immunoglobulin E(IgE)-binding epitopes.Understanding the structural and immunogenic characteristics of allergenic proteins is essential in assessing whether and how food processing techniques reduce allergenicity.We here discuss the impacts of food processing technologies on the modification of physicochemical,structural,and immunogenic properties of allergenic proteins.Detection techniques for characterizing changes in these properties of food allergens are summarized.Food processing helps to reduce allergenicity by aggregating or denaturing proteins,which masks,modifies,or destroys antigenic epitopes,whereas,it cannot eliminate allergenicity completely,and sometimes even improves allergenicity by exposing new epitopes.Moreover,most food processing techniques have been tested on purified food allergens rather than food products due to potential interference of other food components.We provide guidance for further development of processing operations that can decrease the allergenicity of allergenic food proteins without negatively impacting the nutritional profile.

Dietary polyphenols reduced the allergenicity ofβ-lactoglobulin via non-covalent interactions:a study on the structure-allergenicity relationship

Studies showed that complexation of polyphenols with milk allergens reduced their immunogenic potential.However,the relationship between structures of polyphenols and their hypoallergenic effects on milk allergens in association with physiological and conformational changes of the complexes remain unclear.In this study,polyphenols from eight botanical sources were extracted to prepare non-covalent complexes withβ-lactoglobulin(β-LG),a major allergen in milk.The dominant phenolic compounds bound toβ-LG with a diminished allergenicity were identified to investigate their respective role on the structural and allergenic properties ofβ-LG.Extracts from Vaccinium fruits and black soybeans were found to have great inhibitory effects on the IgE-and IgG-binding abilities ofβ-LG.Among the fourteen structure-related phenolic compounds,flavonoids and tannins with larger MWs and multi-hydroxyl substituents,notably rutin,EGCG,and ellagitannins were more potent to elicit changes on the conformational structures ofβ-LG to decrease the allergenicity of complexedβ-LG.Correlation analysis further demonstrated that a destabilized secondary structure and protein depolymerization caused by polyphenol-binding were closely related to the allergenicity property of formed complexes.This study provides insights into the understanding of structure-allergenicity relationship ofβ-LG-polyphenol interactions and would benefit the development of polyphenol-fortified matrices with hypoallergenic potential.

Allergen degradation of bee pollen by lactic acid bacteria fermentation and its alleviatory effects on allergic reactions in BALB/c mice

Food allergy as a global health problem threatens food industry.Bee pollen(BP)is a typical food with allergenic potentials,although it performs various nutritional/pharmacological functions to humans.In this study,lactic acid bacteria(LAB)were used to ferment Brassica napus BP for alleviating its allergenicity.Four novel allergens(glutaredoxin,oleosin-B2,catalase and lipase)were identified with significant decreases in LAB-fermented BP(FBP)than natural BP by proteomics.Meanwhile,metabolomics analysis showed significant increases of 28 characteristic oligopeptides and amino acids in FBP versus BP,indicating the degradation of LAB on allergens.Moreover,FBP showed alleviatory effects in BALB/c mice,which relieved pathological symptoms and lowered production of allergic mediators.Microbial high-throughput sequencing analysis showed that FBP could regulate gut microbiota and metabolism to strengthen immunity,which were closely correlated with the alleviation of allergic reactivity.These findings could contribute to the development and utilization of hypoallergenic BP products.

芝麻过敏原Ses i 2核酸适体的筛选与鉴定研究

芝麻是八大类食物过敏原之一,快速准确识别芝麻过敏原对预防其过敏有重要意义。核酸适配体可以高效识别靶标过敏原,在过敏原检测中有良好的应用前景。为了获得芝麻主要过敏原Ses i 2的特异性核酸适体,本研究以Ses i 2为靶标,通过磁珠筛选法(磁珠-SELEX)开展10轮筛选,经由高通量测序获得6条候补序列(S1~S6),并进行家族性、同源性分析及二级结构预测。结果表明,6条候选核酸适体的重复率可达46.38%,其自由能在-9.02到-2.47 kcal·moL^(-1)之间,根据自由能能量稳定原则,S1和S5吉布斯自由能最低最稳定,分别为-6.70和-9.02 kcal·moL^(-1)。利用ELISA试验进行亲和力测试,结果表明核酸适体S1和S2的亲和能力较强,S1:KD=67.02 nmol·L^(-1),R2=0.925 8,S2:KD=97.65 nmol·L^(-1),R2=0.795 1。核酸适体S1与过敏原Ses i 2的结合力和其他过敏原蛋白相比有显著差异,可视为具有特异性。本研究最终获得一条兼具良好亲和力和特异性的核酸适体S1,为芝麻过敏原快速检测提供了技术支撑。

甲壳类水产品致敏原及特异性免疫制品的研究进展

甲壳类水产品是我国常见的致敏食物之一,除了规避致敏食物的摄入,目前仍无有效的根治方法。因此,致敏原的性质分析及防控技术方法的研究至关重要。本文针对甲壳类水产品致敏原,介绍了致敏原的免疫学特性与分子结构,归纳了致敏原的抗原表位,探讨了应用于致敏原特异性治疗的免疫制品,比较了不同免疫制品的作用机理及优缺点,可为研发水产品过敏的防控技术方法提供参考。