Hydrolyzed Wheat Protein Research Articles

Anaphylactic augmentation by epicutaneous sensitization to acid-hydrolyzed wheat protein in a guinea pig model

Recent reports suggest that hydrolyzed wheat protein (HWP) variants such as Glupearl® 19S (GP19S) induce immediate-type hypersensitivity via epicutaneous (EC) sensitization. The identification of strong allergens is a key step in product assessment before commercial launch. However, few reports have described the estimation of actual and potential anaphylactic sensitizing capacity. In this study we assessed the strength of both the actual and potential anaphylactic sensitizing capacity by investigating the immediate-type hypersensitivity inducing potential of HWP compared with gluten. We assessed these strengths via the EC route using an EC or intradermal (ID) sensitization method. We quantified the strength of immediate-type hypersensitivity by evaluating the titer of serum antibodies isolated from sensitized subjects using passive cutaneous anaphylaxis (PCA) reactions. We also evaluated the cross-reactivity between GP19S and gluten. GP19S and gluten applied by both the sensitization methods induced obvious IgG1-mediated PCA reactions. GP19S had stronger sensitizing potential than gluten, according to the serum titers and dye spot diameters. The difference in antibody titers between GP19S and gluten was 16-fold for the EC method versus 2-fold for the ID method. GP19S cross-reacted with gluten. Acid hydrolysis of gluten increased anaphylactic sensitizing capacity in the EC method. To our knowledge, our study is the first to quantitatively confirm that HWP and gluten can induce immediate-type hypersensitivity through an intact skin. These findings suggest that acid-HWP imposes a higher risk of EC sensitization than gluten because of the ease with which the former confers a sensitizing effect through the intact skin.

Acid hydrolysis of wheat gluten induces formation of new epitopes but does not enhance sensitizing capacity by the oral route: a study in "gluten free" Brown Norway rats.

BackgroundAcid hydrolyzed wheat proteins (HWPs) are used in the food and cosmetic industry as emulsifiers. Cases of severe food allergic reactions caused by HWPs have been reported. Recent data suggest that these reactions are caused by HWPs produced by acid hydrolysis.ObjectivesTo examine the sensitizing capacity of gluten proteins per se when altered by acid or enzymatic hydrolysis relative to unmodified gluten in rats naïve to gluten.MethodsHigh IgE-responder Brown Norway (BN) rats bred on a gluten-free diet were sensitized without the use of adjuvant to three different gluten products (unmodified, acid hydrolyzed and enzymatic hydrolyzed). Rats were sensitized by intraperitoneal (i.p.) immunization three times with 200 µg gluten protein/rat or by oral dosing for 35 days with 0.2, 2 or 20 mg gluten protein/rat/day. Sera were analyzed for specific IgG and IgE and IgG-binding capacity by ELISA. IgE functionality was measured by rat basophilic leukemia (RBL) assay.ResultsRegardless of the route of dosing, all products had sensitizing capacity. When sensitized i.p., all three gluten products induced a strong IgG1 response in all animals. Acid hydrolyzed gluten induced the highest level of specific IgE but with a low functionality. Orally all three gluten products induced specific IgG1 and IgE but with different dose-response relations. Sensitizing rats i.p. or orally with unmodified or enzymatic hydrolyzed gluten induced specific IgG1 responses with similar binding capacity which was different from that of acid hydrolyzed gluten indicating that acid hydrolysis of gluten proteins induces formation of ‘new’ epitopes.ConclusionsIn rats not tolerant to gluten acid hydrolysis of gluten enhances the sensitizing capacity by the i.p. but not by the oral route. In addition, acid hydrolysis induces formation of new epitopes. This is in contrast to the enzymatic hydrolyzed gluten having an epitope pattern similar to unmodified gluten.

Epidemiological link between wheat allergy and exposure to hydrolyzed wheat protein in facial soap.

Recent studies have highlighted the importance of extra-intestinal routes of sensitization to food-related allergens as the cause of epidemics of food allergy. Instances of Japanese women developing food allergy to wheat after exposure to hydrolyzed wheat protein (HWP) present in facial soap have been reported. However, the epidemiologic impact of these ingredients as a cause of food allergy has not been well studied. To clarify the epidemiological relationship between food allergy to wheat and contact exposure to HWP, a case-control study of Japanese women aged 20-54years with self-reported wheat allergy (WA) (cases, n=157) and age-matched control subjects without WA (controls, n=449) was performed using a large-scale Web-based research panel. Subjects answered a Web-based questionnaire regarding the use of skin and hair care products, as well as other possible risk factors. Current use of an HWP-containing facial soap (Cha no Shizuku; Yuka) was significantly associated with an increased risk of WA (adjusted odds ratio, 2.6; 95% confidence interval, 1.2-5.7; frequencies of current use in cases and controls; 11% and 6%, respectively). Use of Cha no Shizuku was more common in subjects with more recent-onset WA, implying that this soap may have contributed to the recent epidemic of WA. An epidemiological relationship between WA and contact exposure to HWP has been documented. This study implicates a possible role of contact exposure to food-derived protein hydrolysates as a risk factor for the development of food allergy manifesting itself as anaphylaxis.

Protein supplements on delayed onset muscle soreness in active men and women (633.1)

The purpose of this study was to compare the effects of different protein supplements on delayed onset muscle soreness (DOMS) in active men and women. Subjects completed four trials (whey protein isolate, WPI; soy protein isolate, SPI; and hydrolyzed wheat protein, HWP; and water, CON) in a double‐blind randomized fashion. During each trial, subjects consumed one of the treatments, rested for 90 minutes, then ran on a treadmill at 75% HRpeak at ‐9 percent grade for 30 min, DOMS was assessed with a tension gauge post‐run and 24 hr after the downhill run. Diet over the 24 hours was designed to be the same for all conditions based on a self‐selected menu. Repeated Measures ANOVA was used to detect differences between treatments with significance set at p < 0.05. Dietary protein elicited a decreased DOMS compared with water, with no differences observed between WPI, SPI, or HWP. These data indicate that dietary protein decreased DOMS when ingested prior to muscle damaging exercise in active men and women.Grant Funding Source: Supported by USDA/AES Hatch funds (KS‐475)

Specific IgE for wheat in tear fluid of patients with allergic conjunctivitis

Context: Allergy to hydrolyzed wheat protein in facial soap has become a major social issue in Japan. It has been reported that the most frequent early symptoms of allergy to hydrolyzed wheat protein in soap are allergic conjunctivitis and rhinitis, while wheat-dependent exercise-induced anaphylaxis can be induced by long-term use.Objective: We evaluated the relation between tear fluid levels of specific IgE for wheat and the features of allergic conjunctivitis.Methods: A prospective, non-randomized, cross-sectional study was conducted in 103 patients with moderate to severe allergic conjunctivitis (allergic group) and 20 age- and sex-matched healthy control subjects (control group). Specific IgE for wheat was measured in tear fluid with an immunochromatography assay, and a skin prick test (SPT) was also performed. Symptoms (sneezing, rhinorrhea, nasal obstruction, ocular itching, and lacrimation) were assessed in each subject along with the activities of daily living (ADL) score and the total ocular symptom score for allergic conjunctivitis. A severity score (0, 1, 2, or 3) was assigned for various changes of the palpebral and bulbar conjunctiva, as well as for limbal and corneal lesions associated with allergic conjunctivitis.Results: The IgE positive rate and specific IgE score were both higher in the allergic group than in the control group (71.8% versus 40.0% and 1.9 ± 0.7 versus 1.4 ± 0.5). A positive SPT for wheat was also more frequent in the allergic group than in the control group (6.8% versus 0.0%). Within the allergic group, patients with a positive SPT had higher specific IgE scores than patients with a negative SPT (3.3 ± 0.5 versus 1.8 ± 0.6, p < 0.001). In the allergic group, the wheat IgE level in tear fluid was correlated with the severity of allergic conjunctivitis symptoms, including ocular itching (r = 0.665), tearing (r = 0.672), and the total ocular symptom score (r = 0.204). Wheat IgE in tear fluid was also correlated with the severity of rhinitis symptoms, including sneezing (r = 0.610), nose blowing (r = 0.640), and nasal obstruction (r = 0.677). Furthermore, the tear fluid wheat IgE score was correlated with five objective features of allergic conjunctivitis (p < 0.05).Conclusions: These results suggest that wheat allergy may be involved in the development of allergic conjunctivitis.

Chronic Headaches and Sleepiness Caused by Facial Soap (Containing Hydrolyzed Wheat Proteins)-Induced Wheat Allergy

A 38-year-old woman was suffering from irregular headaches and sleepiness. She had used soap containing Glupearl 19S (hydrolyzed wheat proteins) every day for approximately one year and had experienced an episode of rash eruption on her face seven months ago. Wheat-specific IgE antibodies were detected in her serum. A Western blot analysis revealed a high titer of IgE antibodies against Glupearl 19S and wheat proteins. The patient was sensitive to these compounds in a skin prick test. After avoiding eating wheat, her headaches and sleepiness disappeared. A hidden food allergy is a possible cause of these symptoms.

Food Allergen in Cosmetics

In Japan, two patients who had been primary sensitized to hydrolyzed wheat protein (HWP) present in facial soap and subsequently experienced wheat-dependent exercise-induced anaphylaxis (WDEIA) after the ingestion of normal wheat products were reported in 2009 as first cases. Since that report, more than 1900 patients with such an allergy have been reported (through June 20, 2013) from various institutes all around Japan. Importantly, the majority of the patients used the same facial soap (Cha no Shizuku(®)) containing acid-hydrolyzed wheat protein (acid-HWP). The commercial acid-HWP contained in the facial soap (Glupearl 19S(®), average molecular weight of 30-50 kDa) was produced from gluten after partial hydrolysis with hydrogen chloride at 95℃ for 40 minutes. In this presentation, I would like to summarize the mechanism of the induction of immediate hypersensitivity by HWP which has been reported by us and other European groups.

A New Reliable Method for Detecting Specific IgE Antibodies in the Patients with Immediate Type Wheat Allergy due to Hydrolyzed Wheat Protein: Correlation of Its Titer and Clinical Severity

Immediate-type wheat allergy caused by a specific hydrolyzed wheat protein (HWP-IWA), Glupearl 19S (GP19S), typically develops food-dependent exercise-induced anaphylaxis (FDEIA), but is different from conventional FDEIA, or simple wheat allergy in many aspects. The skin prick test (SPT) is considered to be the most effective method for diagnosis of HWP-IWA. As SPT is a relatively qualitative method, we developed quantitative and high-throughput test method for HWP-IWA. An enzyme-linked immunosorbent assay (ELISA)-based GP19S-specific IgE assay was tested using sera from 14 HWP-IWA and five conventional wheat-dependent exercise-induced anaphylaxis (CO-WDEIA) patients, as well as five healthy subjects. Then a validation study at five different institutions was carried out using sera from 10 HWP-IWA and five CO-WDEIA patients, as well as five healthy subjects different from the previous studies. The mean unit values converted from measured absorbance of ELISA were 68.3, 1.3 and 1.1 respectively. Furthermore, the validation study revealed reproducible results across all five institutions, with the standard deviation (SD) being 0.3-0.4 for the healthy group, 0.2-0.6 for the CO-WDEIA group, and 3.8-9.6 for HWP-IWA group except for one case. One case of HWP-IWA was excluded from analysis due to the high SD of 53.3 units, indicating that samples with a unit value > 100.0 will affect inter-laboratory reproducibility. Our findings suggest that the ELISA-based GP19S-specific IgE assay can be used to test HWP-IWA using venous blood samples, except for those with a unit value > 100.0.

加水分解コムギアレルギー：最新の知見

加水分解コムギアレルギー：最新の知見

Clinical aspects of 41 cases performed challenge test with immediate-type wheat allergy due to hydrolyzed wheat protein (HWP-IWA)

Immediate-type wheat allergy due to hydrolyzed wheat protein (HWP-IWA) supplemented soap has been a serious social issue. We investigated the significance of challenge test and other tests not only to diagnose HWP-IWA but to know the symptoms of each patients. From January 2010 to June 2012, we performed challenge test in 41 cases with positive prick test of Glupearl 19S®, a major allergic HWP found in Cha no Shizuku®. Thirty nine of 41 cases were challenge test positive. In two patients who showed positive skin prick test but negative for challenge test, titer of specific IgE antibody and/or basophil histamine release test against Glupearl 19S® revealed positive reaction. Challenge test is not included in indispensable examinations for the diagnosis of HWP-IWA in the recommended guidelines. However, challenge test is still a useful tool for assessing actual severity of each patient's symptoms and determining the timing of cessation of wheat avoidance. In addition, combined check-up with several laboratory examination and challenge test may help appropriate diagnosis of HWP-IWA.

EAAS5-2 Remission rate of patients with wheat allergy sensitized to hydrolyzed wheat protein in facial soap(EAAS5 Food allergy and related diseases,Free Paper Session 2,East Asia Allergy Symposium 2014)

EAAS5-2 Remission rate of patients with wheat allergy sensitized to hydrolyzed wheat protein in facial soap(EAAS5 Food allergy and related diseases,Free Paper Session 2,East Asia Allergy Symposium 2014)

Occupational rhinitis, asthma, and contact urticaria caused by hydrolyzed wheat protein in hairdressers

Hydrolyzed wheat protein (HWP) is a common ingredient in food and cosmetic products. Immediate hypersensitivity to HWP has been reported,1,2 and HWP present in foods may cause severe allergic reactions.3 HWP in cosmetic products has been reported to cause allergic contact urticaria (CU)1,4 and even to induce wheat-dependent exercise-induced anaphylaxis (WDEIA) in consumers.5

Tissue transglutaminase generates deamidated epitopes on gluten, increasing reactivity with hydrolyzed wheat protein–sensitized IgE

Tissue transglutaminase generates deamidated epitopes on gluten, increasing reactivity with hydrolyzed wheat protein–sensitized IgE

Enzymatic hydrolysis of gluten proteins changes the dose-response of oral sensitization, but not the gluten epitopes

Background Wheat gluten proteins are very complex proteins containing hundreds of components present as monomers, oligomers and polymers that by definition are not soluble in water. To improve utilization, gluten proteins can be hydrolyzed by enzymes or acid. This increases solubility and provide proteins with new functional properties. During the last decade, cases of severe allergic reaction to hydrolyzed wheat proteins have been reported in subjects tolerant to wheat.

Combining Analyses of Basophil Allergen Threshold Sensitivity, CD-sens, and IgE Antibodies to Hydrolyzed Wheat, ω-5 Gliadin and Timothy Grass Enhances the Prediction of Wheat Challenge Outcome

Background: Wheat is a common food causing allergy which has implications on the quality of life. The diagnosis of IgE-mediated wheat allergy is based on the clinical history and presence of IgE antibodies (IgE-Ab) in skin or blood, and the results of an oral food challenge which is time consuming and associated with risks. An improved diagnostic workup is needed for wheat allergy. The objective was to examine the relationship between wheat challenge, CD-sens and IgE-Ab to related allergens in wheat-allergic children and investigate if a combination of different markers could enhance the prediction of challenge outcome. Method: Twenty-four children (aged 1-15 years) with a wheat allergy diagnosis underwent an open wheat challenge. CD-sens and IgE-Ab to wheat, hydrolyzed wheat protein (HWP), ω-5 gliadin and timothy grass were analyzed and related to the challenge outcome. Results: A positive challenge was seen in 12/24 children. Children reacting to the challenge had higher IgE-Ab concentrations to wheat, ω-5 gliadin and HWP (p < 0.01) and a tendency to higher wheat CD-sens values (p = 0.08) than nonreacting children. Combining wheat CD-sens >150 and IgE-Ab to wheat >20 kU_A/l, or ω-5 gliadin >0.1 kU_A/l predicted the challenge outcome in 83% of the patients. Most children with IgE-Ab to wheat also had IgE-Ab to timothy. Seven of 9 challenge-positive children had a positive CD-sens to HWP and IgE-Ab to HWP >8 kU_A/l. Conclusion: Combining CD-sens and IgE-Ab to wheat or wheat components could be useful in the diagnosis and follow-up of wheat-allergic children.

Immediate contact urticaria caused by hydrolysed wheat proteins in a child via maternal skin contact sensitization

Immediate contact urticaria caused by hydrolysed wheat proteins in a child via maternal skin contact sensitization

Characterization of the Causative Allergens for Wheat-Dependent Exercise-Induced Anaphylaxis Sensitized with Hydrolyzed Wheat Proteins in Facial Soap

Hydrolyzed wheat proteins (HWP) were reported to cause wheat-dependent exercise-induced anaphylaxis (WDEIA) by transcutaneous sensitization through the use of HWP-containing soap in Japan. These patients developed the allergic reactions not only with the use of soap, but also on exercise after the intake of wheat protein (WP). In this study, we identified the allergens and its IgE-binding primary epitopes in WP for HWP-WDEIA patients.

Characterization of Causative Allergens for Wheat-Dependent Exercise-Induced Anaphylaxis Sensitized with Hydrolyzed Wheat Proteins in Facial Soap

In Japan, hydrolyzed wheat proteins (HWP) have been reported to cause wheat-dependent exercise-induced anaphylaxis (WDEIA) by transcutaneous sensitization using HWP-containing soap. Patients develop allergic reactions not only with soap use, but also with exercise after the intake of wheat protein (WP). ω5-Gliadin and HMW-glutenin were identified as major allergens in conventional WP-WDEIA patients. However, the allergens in HWP-WDEIA have yet to be elucidated. Sera were obtained from 22 patients with HWP-sensitized WDEIA. The allergenic activities of HWP and six recombinant wheat gluten proteins, including α/β-, γ-, ω1,2- and ω5-gliadin and low- and high molecular weight (HMW)-glutenins, were characterized by immunoblot analysis and histamine releasing test. IgE-binding epitopes were identified using arrays of overlapping peptides synthesized on SPOTs membrane. Immunoblot analysis showed that IgE antibodies (Abs) from HWP-WDEIA bound to α/β-, γ- and ω1,2-gliadin. Recombinant γ-gliadin induced significant histamine release from basophils in eight of 11 patients with HWP-WDEIA. An IgE-binding epitope "QPQQPFPQ" was identified within the primary sequence of γ-gliadin, and the deamidated peptide containing the "PEEPFP" sequence bound with IgE Abs more strongly compared to the native epitope-peptide. The epitope-peptide inhibited IgE-binding to HWP, indicating that the specific IgE to HWP cross-reacts with γ-gliadin. HWP-WDEIA patients could be sensitized to HWP containing a PEEPFP sequence, and WDEIA symptoms after WP ingestion could partly be induced by γ-gliadin. These findings could be useful to help develop tools for diagnosis and desensitization therapy for HWP-WDEIA.

The Sensitivity and Clinical Course of Patients with Wheat-Dependent Exercise-Induced Anaphylaxis Sensitized to Hydrolyzed Wheat Protein in Facial Soap - Secondary Publication

Recently, an increasing number of patients with wheat-dependent exercise-induced anaphylaxis (WDEIA) have been reported in Japan. Most of them had developed this condition during or after using hydrolyzed wheat protein (HWP)-containing soap (HWP-WDEIA). To clarify the relation between WDEIA and HWP-containing soap and their prognosis, we retrospectively studied the patients who visited Hiroshima University Hospital and were diagnosed as WDEIA from January 2010 to June 2011. We took detailed clinical histories, performed skin prick tests, serum immunoassays for antigen-specific IgE and basophil histamine release test, and followed up their clinical courses after the diagnosis. Among 36 patients with WDEIA, 30 patients had used only one type of HWP-soap. The patients with HWP-WDEIA were mainly women and had developed facial symptoms and angioedema. They suffered from blood pressure reductions less frequently than patients with conventional WDEIA. The levels of gluten-specific IgE were higher than those of omega-5 gliadin in patients with HWP-WDEIA (P < 0.05, One-way ANOVA). All patients with HWP-WDEIA were positive against HWP in histamine release test. Among the conventional wheat antigens, glutenins induced the highest histamine release from basophils of patients with HWP-WDEIA. The sensitivities of patients against glutens and glutenins were reduced over months along with the discontinuance of HWP-soap. The development of HWP-WDEIA is associated with the use of HWP-soap. The sensitivity to HWP that cross reacts with non-processed wheat may be reduced or possibly cured after the discontinuation of HWP-soap.

Higher allergenicity of high molecular weight hydrolysed wheat protein in cosmetics for percutaneous sensitization

Wheat protein derivatives are used in a variety of products worldwide. Gluten is commercially used 'as is' or with modifications such as hydrolysis, which is carried out to overcome its insolubility. Several cases of contact urticaria following exposure to hydrolysed wheat protein (HWP) in cosmetics or of anaphylaxis caused by deamidated gluten in food or non-food products have been described. To evaluate the types of HWP that have higher allergenicity for percutaneous sensitization. We enrolled 7 patients with wheat-dependent exercise-induced anaphylaxis who had been sensitized to HWP primarily through the percutaneous and/or the rhinoconjunctival route by using facial soap containing HWP. Reaction to wheat proteins was confirmed by IgE immunoblotting and basophil CD203c expression with six HWP variants. The IgE of all the patients reacted to HWPs composed of large polypeptide aggregates. High molecular weight (MW) HWPs were also found to induce significant enhancement of basophil CD203c expression. HWPs composed of large polypeptide aggregates possibly induce sensitization to a greater degree than lower-MW HWPs. Basophil surface CD203c expression is useful for evaluating the allergenicity of HWPs.