Citrus Tachibana Leaf Extract Mitigates Symptoms of Food Allergy by Inhibiting Th2‐Associated Responses

Although the incidence of food allergy continues to rise, there have been no effective therapeutic strategies. Citrus fruits contain a number of bioactive flavonoids with immune‐regulatory functions. The objective of this study was to determine whether Citrus tachibana (fruit body with peel, leaves, and branch) can protect against the development of food allergy and the mechanism behind it, and to identify the active compound(s) responsible. We found that C. tachibana leaf extract (CLE) mitigated ovalbumin (OVA)‐induced food allergy symptoms including increased rectal temperature, diarrhea, and anaphylaxis. This mitigation was likely due to CLE‐mediated decreases in cytokine release from T‐helper 2 cells (Th2 cells) in mesenteric lymph nodes. Moreover, higher levels of CLE attenuated systemic Th2 cell–mediated responses in mouse splenocytes sensitized with OVA+Alum. This was evidenced by CLE‐mediated reductions in Th2 cytokine release, including interleukin (IL)‐4, IL‐5, and IL‐13, but not the Th1 cytokines IL‐12 and interferon (IFN)‐γ, which was attributable to decreased gene expression levels. We also identified kaempferol as the most potent compound for reducing Th2‐associated responses in splenocytes. The findings of this study suggest that CLE suppresses Th2‐cell–mediated immune responses, contributing to alleviation of food allergy symptoms, and that kaempferol is a flavonoid with potential antiallergenic activity that targets Th2 cell–induced responses.     

肠道黏膜免疫细胞在食物致敏中的作用研究进展

食物过敏是食品安全的重要问题之一,且大部分食物过敏是在胃肠道消化过程中引发的。食物过敏原蛋白 经胃肠道消化后形成肽段,其中部分消化肽在通过肠上皮组织屏障进入肠道黏膜系统后,被递呈给相关免疫细胞, 从而引发肠道黏膜系统中一系列免疫应答反应。本文阐述了消化肽进入肠道黏膜后,肠上皮细胞、树突状细胞、B 淋巴细胞及CD4＋ T淋巴细胞在黏膜免疫致敏过程中所起的作用,为探讨食物过敏引发胃肠道免疫的工作机制提供 了理论依据。     

Immunomodulating peptides for food allergy prevention and treatment

Among the most promising strategies currently assayed against IgE-mediated allergic diseases stands the possibility of using immunomodulating peptides to induce oral tolerance toward offending food allergens or even to prevent allergic sensitization. This review focuses on the beneficial effects of food derived immunomodulating peptides on food allergy, which can be directly exerted in the intestinal tract or once being absorbed through the intestinal epithelial barrier to interact with immune cells. Food peptides influence intestinal homeostasis by maintaining and reinforcing barrier function or affecting intestinal cell-signalling to nearby immune cells and mucus secretion. In addition, they can stimulate cells of the innate and adaptive immune system while supressing inflammatory responses. Peptides represent an attractive alternative to whole allergens to enhance the safety and efficacy of immunotherapy treatments. The conclusions drawn from curative and preventive experiments in murine models are promising, although there is a need for more pre-clinical studies to further explore the immunomodulating strategy and its mechanisms and for a deeper knowledge of the peptide sequence and structural requirements that determine the immunoregulatory function.     

免疫低下和食物过敏小鼠外周血中细胞因子的变化

建立小鼠免疫低下模型和食物过敏模型,采用ELISA法检测外周血中细胞因子(IL-4、IL-6、IL-10、IL-12、IFN-γ、TGF-β)的变化。结果表明：与空白对照组比较,腹腔注射环磷酰胺所建立免疫低下小鼠胸腺指数和脾脏指数均有显著降低(P≤0.05),且在第3天达到最低;经卵清蛋白致敏小鼠外周血IgE水平极显著升高(P≤0.01)。免疫低下小鼠外周血中IFN-γ(除第3天外)、TGF-β含量(除第3天外)和IFN-γ/IL-4比值均显著降低,而IL-10水平也有下降趋势;除第2天外IL-6水平显著升高,但IL-4水平只在第3天显著性增高,IL-12先是下降随后呈现增多的趋势;食物过敏小鼠的IL-4、IL-6、IL-10和TGF-β水平均有显著性增高,IFN-γ/IL-4比值有显著性下降,而IFN-γ和IL-12没有明显的变化。因此,免疫低下和食物过敏小鼠的Th1/Th2细胞平衡均向Th2细胞偏移,免疫低下对Th1细胞的影响大,食物过敏对Th2细胞的影响更明显。     

Diagnosis and therapy of food allergy

According to the recently revised nomenclature for allergy the term "Food Hypersensitivity" is proposed to define a reaction on food exposure causing objectively reproducible symptoms or signs at a dose tolerated by normal subjects. Those reactions to food in which immunologic mechanisms are demonstrated comprise the term "Food Allergy". Immunologic reactions to food in which an immunoglobulin E (IgE)-mediated mechanism is established are defined as IgE-mediated food allergy. This review focuses on IgE-mediated allergic reactions to foods.     

An adjuvant-free mouse model to evaluate the allergenicity of milk whey protein

Milk allergy is the most common type of food allergy in humans with the potential for fatality. An adjuvant-free mouse model would be highly desirable as a preclinical research tool to develop novel hypoallergenic or nonallergenic milk products. Here we describe an adjuvant-free mouse model of milk allergy that uses transdermal sensitization followed by oral challenge with milk protein. Groups of BALB/c mice were exposed to milk whey protein via a transdermal route, without adjuvant. Systemic IgG1 and IgE antibody responses to transdermal exposure as well as systemic anaphylaxis and hypothermia response to oral protein challenge were studied. Transdermal exposure resulted in a time- and dose-dependent induction of significant IgE and IgG1 antibody responses. Furthermore, oral challenge of sensitized mice resulted in significant clinical symptoms of systemic anaphylaxis within 1 h and significant hypothermia at 30 min postchallenge. To study the underlying mechanism, we examined allergen-driven spleen cell T-helper 2 cytokine (IL-4) responses. There was a robust dose- and time-dependent activation of memory IL-4 responses in allergic mice but not in healthy control mice. These data demonstrate for the first time a novel transdermal sensitization followed by oral challenge mouse model of milk allergy that does not use adjuvant. It is expected that this model may be used not only to study mechanisms of milk allergy, but also to evaluate novel milk products for allergenic potential and aid in the production of hypo- or nonallergenic milk products.     

虾制品致敏性诱发豚鼠相关细胞因子及Th1/Th2细胞平衡的变化

以消减致敏性南美白对虾的虾仁、虾肉和虾蛋白为样品,以pH 7.5的磷酸盐缓冲液为阴性对照,未处理的虾蛋白为阳性对照,建立豚鼠过敏模型,研究过敏豚鼠血清中相关细胞因子与食物过敏的相关性,并推断食物过敏对辅助性T（type 1/type 2 T-helper, Th1/Th2）细胞平衡的影响。收集过敏豚鼠的血清,采用酶联免疫吸附测定法检测血清中免疫球蛋白E（immunoglobulin E,IgE）、组胺（histamine,HIS）、相关细胞因子（白细胞介素（interleukin,IL）-1、IL-2、IL-3、IL-4、IL-6、IL-10、肿瘤坏死因子-α（tumor necrosis factor α,TNF-α）、干扰素-γ（interferon-γ,IFN-γ）质量浓度。结果表明,用未处理的虾蛋白致敏豚鼠,用致敏性消减程度不同的虾制品（虾蛋白、虾肉、虾仁）提取的蛋白激发,消减致敏性的虾蛋白、虾肉和虾仁提取蛋白激发后豚鼠血清中IgE含量分别为（3.905±0.120）、（4.813±0.188）、（5.199±0.327）U/mL,HIS质量浓度分别为（16.437±1.120）、（19.656±1.080）、（21.071±1.732）μg/mL,激发后血清中IgE和HIS质量浓度变化与致敏性程度呈正相关,致敏性越低,血清中IgE和HIS质量浓度越低。过敏豚鼠血清中IL-1、IL-2、IL-3、IL-4、IL-6、TNF-α质量浓度的变化与虾制品致敏程度呈正相关;同时,与IgE和HIS质量浓度变化规律具有一致性;血清中IL-10质量浓度变化与致敏性程度呈负相关;因此,这些细胞因子与食物过敏具有很好的相关性。过敏血清中IFN-γ没有呈现规律性变化,但IFN-γ/IL-4随着致敏性的增强而减小。因此,推测虾制品激发过敏豚鼠的Th1/Th2细胞平衡向Th2细胞偏移。     

肉类过敏研究进展

近年来食物引起的过敏性疾病发病率持续增高,已成为一种严重的食品安全和公共卫生问题。肉类食物作为人类的重要食物来源之一,其市场不断扩大,与此同时,肉类引起的食物过敏也逐渐增加。肉类过敏是IgE介导的Ⅰ型变态反应,发病速度快,可引发多种不同的临床症状,除了食物过敏的基本症状外,还可造成皮肤、呼吸系统疾病。此外,近年来还发现了一类新型肉类过敏疾病,与传统肉类过敏不同,其发病过程表现为延迟发作。本文综述了近年来肉类过敏的研究进展,介绍了几种肉类过敏性疾病及致敏机理,并综述了导致哺乳动物肉类过敏的过敏原以及肉类过敏评估方法。     

Food allergy

Adverse clinical reactions to food associated with disturbed immunologic function (food allergy) affect 1-3% of the population and vary from life-threatening to a minor inconvenience. They must be differentiated from reactions caused by toxins, pharmacologic agents, enzyme deficiences and non-specific release of inflammatory mediator substances. Enteric absorption of food protein antigens which may occur despite an array of gastrointestinal protective mechanisms normally induces both a protective immune response and immunologic tolerance. Quantitative changes in absorption related to deficient protective mechanisms or excessive antigen load may contribute to the development of an allergic immune response and explain the greater incidence of food allergy in infants and children. Important factors include immunologic immaturity, enhanced macromolecular mucosal transport, intrauterine and neonatal malnutrition, breast feeding and infection. Double-blind food challenge tests remain as the most definitive diagnostic yardstick but carefully standardized skin tests may be helpful if interpreted in the context of the clinical history. Despite the association of food allergy with food antigen specific IgE hypersensitivity, immune complex formation and lymphocyte sensitization the pathophysiological changes which result in symptoms remain obscure. Recent advances have clarified many aspects of our knowledge of food allergy but inevitably have raised many more questions for future study.     

Prophylactic effect of Lactobacillus oral vaccine expressing a Japanese cedar pollen allergen

Lactic acid bacteria (LAB) represent an attractive delivery vehicle for oral allergy vaccine because of their safety as a food microorganism as well as their potent adjuvant activity triggering anti-allergic immune response. Here, we report the generation of recombinant LAB expressing a major Japanese cedar pollen allergen Cry j 1 (Cry j 1-LAB), and their prophylactic effect in vivo. To facilitate heterologous expression, the codon usage in the Cry j 1 gene was optimized for the host LAB strain Lactobacillus plantarum by the recursive PCR-based exhaustive site-directed mutagenesis. Use of the codon-optimized Cry j 1 cDNA and a lactate dehydrogenase gene fusion system led to a successful production of recombinant Cry j 1 in L. plantarum NCL21. We also found that oral vaccination with the Cry j 1-LAB suppressed allergen-specific IgE response and nasal symptoms in a murine model of cedar pollinosis.     

T细胞及其表位在食物过敏中的研究进展

食物过敏蛋白经胃肠道消化后经树突状细胞递呈给T细胞后,会启动过敏反应的发生。在此过程中,不同T细胞发挥了不同的作用,其中Th1/Th2下调被认为是引发过敏反应的主要工作机制,Th9、Th17和Tfh等细胞在过敏反应发展中的作用不容忽视。作者介绍了不同T淋巴细胞的作用,探讨了T细胞表位预测及定位以及目前T细胞表位在常见过敏原蛋白中的应用,综述了T细胞在食物过敏中的研究进展。     

Single nucleotide polymorphisms (SNPs) in key cytokines may modulate food allergy phenotypes

Single nucleotide polymorphisms (SNPs) can play a direct or indirect role in phenotypic expression in food allergy pathogenesis. Our goal was to quantitate the expression of SNPs in relevant cytokines that were expressed in food allergic patients. SNPs in cytokine genes IL-4 and IL-10 are known to be important in IgE generation and regulation. We examined IL-4 (C-590T), IL-4Rα (1652A/G) and IL-10 (C-627A) SNPs using real-time PCR followed by restriction fragment length polymorphism (RFLP) analysis. Our results show that the AA, AG and GG genotypes for IL-4Rα (1652A/G) polymorphisms were statistically different in radioallergosorbent test (RAST) positive versus negative patients, and although no statistically significant differences were observed between genotypes in the IL-4 (C-590T) and IL-10 (C-627A) SNPs, we observed a significant decrease in IL-4 (C-590T) gene expression and increase in IL-4Rα (1652A/G) and IL-10 (C-627A) gene expression between RAST⁺ versus RAST^? patients, respectively. We also observed significant modulation in the protein expression of IL-4 and IL-10 in the serum samples of the RAST⁺ patients as compared to the RAST^? patients indicating that changes in SNP expression resulted in altered phenotypic response in these patients.     

Wheat Allergy and Intolerence; Recent Updates and Perspectives

The current review paper highlights the complicacies associated with communities relying on wheat as their dietary staple. Although, wheat is an important source of nutrients but is also linked with allergenic responses in genetically susceptible subjects. The wheat proteins especially α-amylase inhibitors, ω-5 gliadins, prolamins, nonprolamin, glucoprotein, and profilins are of significance importance. The allergenic responses are further categorized into IgE-mediated and non-IgE-mediated reactions. Conjugation and degranulation of the IgEs with the allergens results in release of several mediators. In contrary, non-IgE-mediated wheat allergy depends on immune complexes formed by food and food antibodies and cell-mediated immunity. As results, different diseases tend to occur on the completion of these reactions, i.e., celiac disease, baker's asthma, diarrhea, atopic dermatitis, and urticaria. This instant paper highlighted the concept of food allergy with special reference to wheat. The models are developed that are included in this paper showing the wheat allergen, their possible routes, impact on human health, and indeed possible remedies. The paper would provide the basic information for the researchers, common man, and allied stakeholders to cater the issue in details. However, the issue needs the attention of the researchers as there is a need to clarify the issues of wheat allergy and wheat intolerance.     

Development of a Biological Assay to Determine the Allergenic Potential of Foods

Food allergies represent a risk for many people in industrialized countries. Unrecognizable allergenic proteins of foodstuffs may be present as ingredients that are not labeled or as unknown cross-contamination. Such hidden allergens can cause severe reactions in allergics, even at minute quantities, sometimes with fatal outcome. For the verification of the presence of allergenic food constituents, analytical methods such as ELISA and PCR have been developed. However, these tests cannot measure allergenic potential. For this reason, a test system that measures the biological activity of allergens has been developed. It is based on the cellular mechanisms of the type I allergy. Rat basophilic leukemia cells (RBL-2H3) were transfected with the genes of the human high affinity receptor for IgE. The resulting cell line expressed the human receptor α-chain and could bind allergenspecific IgE from allergic subjects, in contrast to the parent cell line. After cross-linking of receptor-bound, allergen-specific human IgE by allergens, the cells released measurable inflammatory mediators. These cells were used for the analysis of a variety of allergen extracts, including extracts prepared from foods containing allergenic hazelnut and peanut. The comparative validation with existing ELISA and PCR for hazelnut and peanut demonstrated similar sensitivity and specificity. The established cell line will be a novel tool in the detection of allergens in complex mixtures, especially to address the issue of their allergenic potential, which cannot be accomplished by classical analytical methods. This will add valuable information about the allergenic potential of food constituents to the risk assessment of foods.     

对虾原肌球蛋白致敏小鼠模型的构建及乳酸菌肠黏膜免疫效应对致敏性的影响

以原肌球蛋白（tropomyosin,TM）为主要过敏原,腹腔注射6 周龄BALB/c雌、雄小鼠,从第14天开始给治疗组小鼠口服灌胃长双歧杆菌婴儿亚种（Bifidobacterium longum subsp.）1.2202或芽孢乳酸菌09.712（Bacillus coagulans 09.712）,连续灌胃20 d。根据小鼠腹泻情况、过敏症状评分、血清中组胺（histamine,HIS）质量浓度、TM特异性免疫球蛋白（immunoglobulin,Ig）E水平变化构建TM致敏及益生菌治疗小鼠模型;通过氯乙酸萘酚酯酶染色、高碘酸希夫试剂特殊染色法检测各组小鼠肠道部位的肥大细胞和杯状细胞,通过免疫组化法检测各组小鼠肠道部位的CD4＋ T细胞和嗜酸性粒细胞;探究肠黏膜免疫在TM致敏及益生菌缓解TM致敏作用中的机理。结果表明：雌性小鼠更适用于构建TM致敏模型;益生菌灌胃可以通过降低血清中HIS质量浓度和IgE水平缓解肠黏膜免疫反应,减轻TM过敏症状。本研究从肠黏膜免疫反应角度探究了其在TM致敏中的作用,为食物过敏的研究提供参考。     

肠道菌群调控下的食物过敏机制研究进展

随着过敏人群数量的快速增加,食物过敏日益受到人们的关注。肠道微生物在机体免疫系统的形成和发展 中扮演着重要角色,越来越多的研究表明,抗生素的使用、膳食结构的改变等因素所引起的肠道菌群紊乱将会增加 人群患食物过敏的机率。本文将从细胞和分子水平综述目前对口服耐受的产生及因其缺失而引起食物过敏的相关机 理的研究,并在此基础上讨论肠道微生物调节食物过敏的潜在通路,最后概述有关益生菌缓解食物过敏的应用及机 制研究。通过了解肠道微生物与宿主黏膜系统的相互作用,可为预防及治疗食物过敏提供新的调控靶点和研究思 路,并为益生菌缓解食物过敏提供理论依据和改进方案,从而减少食物过敏的发生。     

Oral tolerance induction does not resolve gastrointestinal inflammation in a mouse model of food allergy

Scope : Oral immunotherapy (OIT) involving continuous oral administration of allergenic foods has gained attention as a therapy for food allergies. To study the influence of oral administration of allergenic foods on gastrointestinal symptoms including inflammation, we established a mouse model of food‐induced intestinal allergy. Methods and results : BALB/c mice were fed an egg white (EW) diet containing ovalbumin (OVA, a major EW allergen) after intraperitoneal sensitisation with OVA and Alum. The mice on the EW diet for one wk presented gastrointestinal symptoms (i.e. weight loss and soft stools) and inflammation in the small intestines (i.e. duodenum, jejunum and ileum). Further continuous EW diet resolved the weight loss but not the soft stools. Splenic CD4⁺ T‐cells of EW diet‐fed mice on the continuous diet showed less proliferation and cytokine production compared with those of control mice, suggesting tolerance induction by the diet. The continuous EW diet reduced levels of OVA‐specific IgE antibodies, but significantly aggravated the inflammation in the jejunum. Conclusion : Our mouse model would be useful to investigate inflammatory and regulatory mechanisms in food‐induced intestinal allergies. Our results suggest potential gastrointestinal inflammation in patients undergoing OIT as continuous administration of allergenic foods, even though the therapy may induce clinical tolerance.     

Impact of dietary factors and food processing on food allergy

Allergic reactions to food can significantly reduce the quality of life and even result in life‐threatening complications. In addition, the prevalence of food allergy has increased in the last decades in industrialized countries and the mechanisms underlying (increased) sensitization are still not fully understood. It is believed that the development and maintenance of oral tolerance to food antigens is a process actively mediated by the immune system and that this reaction is essential to inhibit sensitization. Ongoing research indicates that different dietary factors also may contribute to immune homeostasis and oral tolerance to food and that food processing modulates allergenicity. One of the major questions in food allergy research is therefore which impact nutrition and food processing may have on allergenicity of food and perhaps on sensitization. We summarize in this review the different dietary factors that are believed to contribute to induction of oral tolerance and discuss the underlying mechanisms. In addition, the functional consequences of allergen modification will be emphasized in the second part as severity of allergic reactions and perhaps sensitization to food is influenced by structural modifications of food allergens.     

儿童食物过敏及其饮食干预治疗研究进展

食物过敏是指由人体免疫系统介导的食物不良反应,常见皮肤、消化道、呼吸道症状,如特应性鼻炎等疾病,严重会出现休克等现象。儿童作为食物过敏易感人群,其生长发育极易受到影响,并且近年来全球儿童食物过敏人群逐年增长,因此对于儿童食物过敏进行预防和治疗有着十分重要的意义。但目前关于儿童食物过敏的流行病学资料仍然比较缺乏,通过查阅中英文文献,本文对儿童食物过敏的常见致敏食物、过敏机制、发病因素及诊治方法进行综述,为食物过敏的预防和治疗方向提供一定的理论依据。