Oral administration of ovalbumin after sensitization attenuates symptoms in a mouse model of food allergic enteropathy

Oral immunotherapy (OIT) is a promising treatment of food allergy. To administer an appropriate oral dose of an allergenic component as OIT to individuals sensitized with a food allergen may prevent inducing food allergic inflammation in them. So we attempted to establish a mouse model to evaluate efficacy for oral administration of food allergen after sensitization. In BALB/c mice sensitized by injecting ovalbumin (OVA) with alum twice, OVA was administered before inducing inflammation by feeding the mice with egg white (EW) diet. Severe inflammatory responses, such as enteropathy, weight loss, IL-4 production, and increase of IgE antibody levels, were suppressed by administration with 4 mg of OVA 7 times before feeding EW diet. OVA administration alone induced a slight Th2 response, but no symptoms. The current study demonstrated that severe food allergic enteropathy could be prevented by pre-administration with appropriate dose of OVA to sensitized mice.     

Experimental food allergy leads to adipose tissue inflammation, systemic metabolic alterations and weight loss in mice

To investigate the consequences of food allergy in adipose tissue and metabolism, we used a murine model in which mice have been sensitized subcutaneously with ovalbumin and further received antigen-containing diet. Allergic mice presented a significant weight loss 7 days after oral challenge with a concomitant decrease in epididymal adipose tissue mass. This decrease was associated with increased lipolysis and local inflammation. In adipose tissue of allergic mice there were increased leukocyte rolling and adhesion in the microvasculature, increased number of leukocytes in the tissue, especially macrophages (F4/80⁺ cells) and increased pro-inflammatory cytokines levels, including TNF-α, IL-6 and CCL2. In addition, we observed low serum concentrations of triglyceride, glucose, total cholesterol and free fatty acids in the allergic mice. Our results suggest that the induction of food allergy in mice leads to adipose tissue inflammation and systemic metabolic alterations that contribute to the weight loss observed.     

Role of IL-4 in aversion induced by food allergy in mice

To ascertain the role of IL-4 in aversion to antigen induced by food allergy, wild type and IL-4 deficient BALB/c mice were sensitized with ovalbumin and challenged orally with egg white. Sensitized wild type mice had increased production of IL-4 by spleen and mesenteric lymph node cells in vitro, higher levels of serum anti-ovalbumin IgE and IgG1, aversion to ingestion of the antigen and loss of body weight after continuous oral challenge. Intestinal changes in wild type sensitized mice included eosinophil infiltration and increased mucus production. The IL-4 deficiency impaired the development of food allergy and the aversion to antigen, suggesting the involvement of the antigen specific antibodies. When IL-4 deficient mice received serum from sensitized wild type donors, the aversion was restored. These results indicate that production of IL-4 and specific IgE/IgG1 antibodies correlate with aversion to antigen induced by food allergy in mice.     

Novel T-cell epitopes of ovalbumin in BALB/c mouse: Potential for peptide-immunotherapy

The identification of food allergen T-cell epitopes provides a platform for the development of novel immunotherapies. Despite extensive knowledge of the physicochemical properties of hen ovalbumin (OVA), a major egg allergen, the complete T-cell epitope map of OVA has surprisingly not been defined in the commonly used BALB/c mouse model. In this study, spleen cells obtained from OVA-sensitized mice were incubated in the presence of 12-mer overlapping synthetic peptides, constructed using the SPOTS^® synthesis method. Proliferative activity was assessed by 72-h in vitro assays with use of the tetrazolium salt WST-1 and led to identification of four mitogenic sequences, i.e., A39R50, S147R158, K263E274, and A329E340. ELISA analyses of interferon (IFN)-γ and interleukin (IL)-4 productions in cell culture supernatants upon stimulation with increasing concentrations of peptides confirmed their immunogenicity.Knowledge of the complete T-cell epitope map of OVA opens the way to a number of experimental investigations, including the exploration of peptide-based immunotherapy.     

食物过敏儿童肠道正常菌群的定量调查与分析

目的探讨儿童食物过敏与肠道正常菌群的关系。方法采集食物过敏儿童与正常儿童粪便,每例作需氧或厌氧培养、计数与鉴定。结果食物过敏儿童肠道中的双歧杆菌、乳酸杆菌、肠杆菌数量与正常儿童差异有显著性（P〈0．05）。结论食物过敏与肠道菌群失调有密切关系。     

Nitration of the egg-allergen ovalbumin enhances protein allergenicity but reduces the risk for oral sensitization in a murine model of food allergy

Background

Nitration of proteins on tyrosine residues, which can occur due to polluted air under “summer smog” conditions, has been shown to increase the allergic potential of allergens. Since nitration of tyrosine residues is also observed during inflammatory responses, this modification could directly influence protein immunogenicity and might therefore contribute to food allergy induction. In the current study we have analyzed the impact of protein nitration on sensitization via the oral route.

Methodology/Principal Findings

BALB/c mice were immunized intragastrically by feeding untreated ovalbumin (OVA), sham-nitrated ovalbumin (snOVA) or nitrated ovalbumin (nOVA) with or without concomitant acid-suppression. To analyze the impact of the sensitization route, the allergens were also injected intraperitoneally. Animals being fed OVA or snOVA under acid-suppressive medication developed significantly elevated levels of IgE, and increased titers of specific IgG1 and IgG2a antibodies. Interestingly, oral immunizations of nOVA under anti-acid treatment did not result in IgG and IgE formation. In contrast, intraperitoneal immunization induced high levels of OVA specific IgE, which were significantly increased in the group that received nOVA by injection. Furthermore, nOVA triggered significantly enhanced mediator release from RBL cells passively sensitized with sera from allergic mice. Gastric digestion experiments demonstrated protein nitration to interfere with protein stability as nOVA was easily degraded, whereas OVA and snOVA remained stable up to 120 min. Additionally, HPLC-chip-MS/MS analysis showed that one tyrosine residue (Y₁₀₇) being very efficiently nitrated is part of an ovalbumin epitope recognized exclusively after oral sensitization.

Conclusions/Significance

These data indicated that despite the enhanced triggering capacity in existing allergy, nitration of OVA may be associated with a reduced de novo sensitizing capability via the oral route due to enhanced protein digestibility and/or changes in antibody epitopes.     

食物过敏婴儿和健康儿肠道菌群分析

目的检测食物过敏婴儿与健康婴儿的大便菌群,为食物过敏与肠道菌群关系的研究提供依据。方法以在重庆儿童医院体检的52例食物过敏婴儿和年龄、性别匹配的100例健康婴儿为研究对象,收集所有对象的新鲜粪便,采用直接快速涂片法分析肠道菌群。结果食物过敏婴儿革兰阳性杆菌比例降低,革兰阴性杆菌、革兰阳性球菌比例增高[(71.43±8.70)%vs(78.91±9.25)%,(15.69±6.26)%vs(11.85±6.66)%,(11.67±4.02)%vs(7.60±4.27)%,P均<0.001]。不同喂养方式下,食物过敏婴儿的大便菌群比例显示不同的改变,但革兰阳性杆菌比例降低是其共同的特点(P均<0.05)。结论食物过敏婴儿肠道菌群与健康婴儿的肠道菌群差异存在显著性。不同喂养方式的婴儿显示不同的大便菌群组成。肠道正常菌群的改变可能在食物过敏的发生中起一定作用。     

Biotechnologies in new high-throughput food allergy tests: why we need them

The increase in prevalence of food allergies generates a need for more accurate and reliable quantitative allergy testing in order to help diagnosis. In this short review, we briefly outline the history of food allergy testing and extend our comments to current multiplex techniques. Particular emphasis is given to new developments in the protein microarray area, where the use of recent advances in biotechnology has the potential to produce high-throughput devices with improved clinical significance.     

The two faces of mast cells in food allergy and allergic asthma: The possible concept of Yin Yang

The purpose of this review is to discuss the role of mast cells in allergic inflammation. We have focused on inflammation associated with allergic asthma and food allergy. Mast cells are ‘first line of defense’ innate/adaptive immune cells and are widely distributed in tissues in surfaces exposed to the environment. Especially in allergic settings mast cells are extensively studied, as they can be activated to release a wide range of mediators by allergen-IgE specific triggers. In addition, in allergic inflammation mast cells can also be activated non-allergic triggers. Recent studies revealed that mast cells, besides the classical role of pro-inflammatory effector cell, have also emerged as modulators of allergic sensitization and down-regulators of allergic inflammation. Therefore, mast cells can be regarded as ‘Ying Yan’ modulators in allergic responses in intestinal tract and airways. This article is part of a Special Issue entitled: Mast Cells in Inflammation.     

宫内节育器与宫腔支撑球囊联合应用治疗中重度宫腔粘连中的临床效果

摘要 目的：探讨宫内节育器与宫腔支撑球囊联合应用治疗中重度宫腔粘连的临床效果。方法：选择2016年12月至2019年8月于西安交通大学第一附属医院宫腔镜诊疗中心接受宫腔镜下中重度宫腔粘连分解术的患者共96例,采用随机对照表法将其分为两组。对照组48例,术后宫腔内放置宫内节育器(intrauterine contraceptive device,IUD);研究组48例,术后放置宫腔支撑球囊(intrauterine support balloon,ISB),3~5 d后更换为宫内节育器。两组术后均给予为期3月的人工周期治疗。术后1月,复查宫腔镜了解宫腔有无再粘连并取出宫内节育器,以此评估手术效果是否有效,术后3月时随访其月经来潮情况。统计比较两组术后5 d内阴道出血量、C反应蛋白水平、发热、下腹痛以及节育器或支持球囊有无脱落等情况,术后1月手术效果及术后3月月经恢复情况。结果：研究组的手术有效率及月经改善比例均显著高于对照组(P<0.05),术后5 d内阴道出血量显著少于对照组(P<0.05);下腹痛及球囊脱落事件发生率显著低于对照组(P<0.05)。两组在C反应蛋白水平、发热发生率对比无统计学差异(P>0.05)。结论：与单独使用宫内节育器相比,联合使用宫腔支撑球囊能提高中重度宫腔粘连分解手术有效率,改善月经,减少术后出血,不增加感染风险,但球囊易于脱落且会增加患者下腹疼痛。     

Downregulation in aquaporin 4 and aquaporin 8 expression of the colon associated with the induction of allergic diarrhea in a mouse model of food allergy

Food allergies have become increasingly prevalent during the past few decades. Diarrhea is one of the most frequent intestinal symptoms caused by food allergens and is characterized by imbalanced ion exchange and water transfer; however, the underlying mechanism of allergic diarrhea remains unclear. Water transfer across the intestinal epithelial membrane seems to occur via aquaporins (AQPs). However, the molecular mechanism of water transfer and the pathophysiological roles of aquaporins in the intestine have not been fully established. The present studies have focused on the alterations of AQPs in a mouse model of allergic diarrhea in which BALB/c mice developed diarrhea following repeated challenges of orally administered ovalbumin. Quantitative real-time PCR analysis and immunohistochemical technique were used for expression of mRNA and protein of AQPs, respectively. AQP4 and AQP8 mRNA levels were significantly decreased in the proximal colon of allergic mice compared to controls; likewise, expression of AQP4 and AQP8 proteins was reduced in the proximal colon of the allergic mice. These results suggest that allergic diarrhea is associated with a downregulation in AQP4 and AQP8 expression.     

The role of commensal bacteria in the regulation of sensitization to food allergens

The prevalence of life-threatening anaphylactic responses to food is rising at an alarming rate. The emerging role of the gut microbiota in regulating food allergen sensitization may help explain this trend. The mechanisms by which commensal bacteria influence sensitization to dietary antigens are only beginning to be explored. We have found that a population of mucosa-associated commensal anaerobes prevents food allergen sensitization by promoting an IL-22-dependent barrier protective immune response that limits the access of food allergens to the systemic circulation. This early response is followed by an adaptive immune response mediated in part by an expansion of Foxp3⁺ Tregs that fortifies the tolerogenic milieu needed to maintain non-responsiveness to food. Bacterial metabolites, such as short-chain fatty acids, may contribute to the process through their ability to promote Foxp3⁺ Treg differentiation. This work suggests that environmentally induced alterations of the gut microbiota offset the regulatory signals conferred by protective bacterial species to promote aberrant responses to food. Our research presents exciting new possibilities for preventing and treating food allergies based on interventions that modulate the composition of the gut microbiota.     

Identification of the 7S and 11S globulins as percutaneously sensitizing soybean allergens as demonstrated through epidermal application of crude soybean extract

Cutaneous exposure to food allergens can predispose individuals to food allergies. Soybean, a major allergenic food, is an ingredient in various cosmetic products. However, the types of soybean proteins that are percutaneously sensitizing in humans or animal models remain unknown. In this study, BALB/c mice were dorsally shaved and epicutaneously exposed to a crude soybean extract including sodium dodecyl sulfate or distilled water alone. Specific IgEs secreted in response to 7S globulin (Gly m 5), 11S globulin (Gly m 6), Gly m 3, and Gly m 4 were measured using enzyme-linked immunosorbent assays or immunoblots. Exposure to soybean extract elicited the secretion of soybean-specific IgEs. Of the soybean proteins, 7S and 11S globulins acted as percutaneous sensitizers in 6/9 mice (67%). Additionally, IgE bound specifically and preferentially to the 7S globulin β subunit. In conclusion, this is the first report to identify percutaneously sensitizing soybean allergens in a mouse model.     

Air cleaners can reduce the risk of allergic sensitization

Exposure to house-dust mite allergens is a factor in the development of allergic symptoms in atopic patients. Several measures can be proposed to control indoor allergen levels, inducing a clinical benefit. The use of an air cleaner is one simple way of achieving this goal. We conducted a simulation trial in a proper room, to verify the usefulness of a domestic cleaner, based on mechanical air filtration, to reduce the levels of environmental allergens. We checked the presence of mite components by different methods (Aclotest and Der p 1 ELISA), in dust recovered before and after using the air cleaner. Our results indicate that this approach could be useful in significantly lowering the levels of mite allergens.     

食物过敏患儿肠道菌群特征及其与外周血Treg/Th17的相关性

分析食物过敏患儿肠道菌群分布与外周血免疫细胞Treg/Th17的关系。

选择2020年4月至2022年5月本院儿科收治的86例食物过敏患儿作为观察组,另选择同期80例健康儿童作为健康对照组。采集两组儿童新鲜粪便和外周血样本,分别测定肠道菌群数量以及外周血Treg/Th17细胞、血清相关细胞因子水平,并经Pearson相关性分析肠道菌群与外周血Treg/Th17的相关性。

观察组患儿肠道内乳杆菌、双歧杆菌数量均低于健康对照组（均P＜0.05）;观察组患儿外周血内Treg细胞水平、Treg/Th17比值均低于健康对照组,但Th17细胞水平高于健康对照组（均P＜0.05）;观察组患儿的血清白细胞介素-17（interleukin-17,IL-17）水平高于健康对照组,血清转化生长因子-β1（transforming growth factor-β1,TGF-β1）指标低于健康对照组（均P＜0.05）;Pearson相关性分析结果显示,乳杆菌、双歧杆菌与外周血Treg细胞、TGF-β1水平均呈正相关,而与外周血Th17细胞、IL-17水平均呈负相关（均P＜0.05）。

食物过敏患儿伴显著的肠道菌群紊乱与免疫功能异常状况,肠道内乳杆菌和双歧杆菌水平以及外周血Treg细胞、TGF-β1表达减少,外周血Th17细胞、IL-17表达增多。临床应改善患儿肠道微生态环境,调控肠道菌群结构,促进肠道菌群分布平衡恢复。

     

儿童肠道菌群与食物过敏关系的研究进展

食物过敏(food allergy, FA)在儿童中的发生率逐年升高,严重影响其生活质量,已经成为全球面临的公共卫生问题之一。近年来,人们发现FA儿童的肠道菌群组成与健康儿童有显著差异。深入研究发现,肠道菌群可通过调节树突状细胞、辅助性T细胞、调节性T细胞、肥大细胞和粒细胞等免疫细胞维持免疫平衡,也可通过多种方式增强肠道屏障功能,抑制FA的发生。在已有研究基础上,益生菌和益生元在治疗儿童FA方面也得到了一定应用,但目前的应用效果并不明确。本文以婴幼儿FA在全球范围内患者规模日益扩大为背景,综述了肠道菌群影响FA的部分机制,总结了近年部分益生菌和益生元在治疗和预防婴幼儿FA方面的应用,并为肠道菌群在FA发生和发展中的作用机制研究和益生菌及其相关代谢产物在儿童FA治疗和预防中的应用提出了新思路,对促进婴幼儿FA治疗方法和策略的研究有重要意义。     

Cellular and molecular mechanisms of vitamin D in food allergy

Food allergies are becoming increasingly prevalent, especially in young children. Epidemiological evidence from the past decade suggests a role of vitamin D in food allergy pathogenesis. Links have been made between variations in sunlight exposure, latitude, birth season and vitamin D status with food allergy risk. Despite the heightened interest in vitamin D in food allergies, it remains unclear by which exact mechanism(s) it acts. An understanding of the roles vitamin D plays within the immune system at the cellular and genetic levels, as well as the interplay between the microbiome and vitamin D, will provide insight into the importance of the vitamin in food allergies. Here, we discuss the effect of vitamin D on immune cell maturation, differentiation and function; microbiome; genetic and epigenetic regulation (eg DNA methylation); and how these processes are implicated in food allergies.     

A preliminary study of gut dysbiosis in children with food allergy

Gut microbiota of food allergic children was analyzed by high throughput 16S rRNA gene sequencing. Signs of gut dysbiosis, which is likely associated with gut inflammation, was observed in children with food allergies. For example, decreased abundance of genus Akkermansia but increased abundance of Veillonella was found in children with food allergy in comparison with healthy control children.     

PYY(3-36) into the arcuate nucleus inhibits food deprivation-induced increases in food hoarding and intake

Central administration of neuropeptide Y (NPY) increases food intake in laboratory rats and mice, as well as food foraging and hoarding in Siberian hamsters. The NPY-Y1 and Y5 receptors (Rs) within the hypothalamus appear sufficient to account for these increases in ingestive behaviors. Stimulation of NPY-Y2Rs in the Arcuate nucleus (Arc) has an anorexigenic effect as shown by central or peripheral administration of its natural ligand peptide YY (3-36) and pharmacological NPY-Y2R antagonism by BIIE0246 increases food intake. Both effects on food intake by NPY-Y2R agonism and antagonism are relatively short-lived lasting ∼4 h. The role of NPY-Y2Rs in appetitive ingestive behaviors (food foraging/hoarding) is untested, however. Therefore, Siberians hamsters, a natural food hoarder, were housed in a semi-natural burrow/foraging system that had (a) foraging requirement (10 revolutions/pellet), no free food (true foraging group), (b) no running wheel access, free food (general malaise control) or (c) running wheel access, free food (exercise control). We microinjected BIIE0246 (antagonist) and PYY_(3-36) (agonist) into the Arc to test the role of NPY-Y2Rs there on ingestive behaviors. Food foraging, hoarding, and intake were not affected by Arc BIIE0246 microinjection in fed hamsters 1, 2, 4, and 24 h post injection. Stimulation of NPY-Y2Rs by PYY_(3-36) inhibited food intake at 0–1 and 1–2 h and food hoarding at 1–2 h without causing general malaise or affecting foraging. Collectively, these results implicate a sufficiency, but not necessity, of the Arc NPY-Y2R in the inhibition of food intake and food hoarding by Siberian hamsters.     

Immunosuppressive activity of the ethanol extract of Siegesbeckia orientalis on the immune responses to ovalbumin in mice

The in vitro and in vivo immunosuppressive activity of the ethanol extract of Siegesbeckia orientalis (EESO) was studied on the immune responses in mice. EESO significantly suppressed concanavalin A (Con A)- and lipopolysaccharide (LPS)-stimulated splenocyte proliferation in vitro in a concentration-dependent manner. ICR Mice were immunized subcutaneously with ovalbumin (OVA) on days 0 and 14. Beginning on the day of immunization, the mice were administered intraperitoneally with EESO at a single dose of 0.25, 0.5, and 1.0 mg at intervals of 7 days. On day 28, OVA-specific antibodies in serum, and mitogen- and OVA-induced splenocyte proliferation were measured. EESO significantly suppressed Con A-, LPS- and OVA-induced splenocyte proliferation in the OVA-immunized mice in a dose-dependent manner. The OVA-specific serum IgG, IgG1, and IgG2b levels in the OVA-immunized mice were also significantly reduced by EESO. Moreover, reducing effect on the IgG1 antibody of EESO at the dose of 1.0 mg was more significant than that of cyclosporin A (CsA; positive drug). The results suggest that EESO could suppress the cellular and humoral response to ovalbumin in mice, and deserve further investigations to be developed as immunosuppressant.