By now, the digestive stability experiments provided by most authoritative organizations are in vitro tests. Evaluating the protein digestive stability with in vivo models should be more objective. The present study aimed to verify the in vivo digestibility of soybean β-conglycinin β-subunit in Wuzhishan (WZS) minipigs. Three minipigs were surgically fitted with O-stomach and T-ileum cannulae and fed with soybean meals. According to SDS-PAGE, the 50 kD fraction of soybean β-conglycinin β-subunit persisted in the gastric fluid until 6 h after feeding, which was detected at 3 h and clearly visible at 4-6 h in the intestinal fluid. Western blot with anti-β-conglycinin β-subunit McAb confirmed it.
Animals ; Antigens, Plant ; chemistry ; metabolism ; Digestion ; physiology ; Globulins ; chemistry ; metabolism ; Male ; Protein Subunits ; chemistry ; metabolism ; Seed Storage Proteins ; chemistry ; metabolism ; Soybean Proteins ; chemistry ; metabolism ; Swine ; Swine, Miniature ; physiology

OBJECTIVETo establish WZS miniature swine model of beta-conglycinin (7 S) allergy for evaluating the potential allergenicity of genetically modified food.

METHODSTwelve 45-day-old WZS miniature swines from three litters were randomly divided into three groups (control group; 4% 7 S group and 8% 7 S group, n = 4), which were respectively gastric sensitized (day 0 - 10) and oral challenged (day 6 - 18, 31) to induce anaphylactic reactions. Clinical symptoms, skin prick reactions were recorded. At day 10, 19 and 32, serum IgG, IgE, histamine and cytokines levels were measured by ELISA.

RESULTSDiarrhea at different degrees were observed in 4% and 8% 7 S groups. The skin erythema reactions in grade "-", "+/-", "+", "++" of control group respectively were 2/4, 2/4, 0/4, 0/4, of 4% 7 S group were 0/4, 0/4, 2/4, 2/4 and of 8% 7 S group were 0/4, 0/4, 1/4, 3/4. The serum IgE and histamine levels of day 11, 19 and 32 were all significantly and positively correlated (Pearson coefficients = 1, P = 0.000). The serum IgG, IgE and histamine levels all reached the peak after 7 S groups were oral challenged at day 19.Compared with the control group, serum IgG (lg IgG: 2.95 +/- 0.31 vs 2.29 +/- 0.25, t = 3.19, P = 0.011), IgE (lg IgE: 2.45 +/- 0.30 vs 1.77 +/- 0.23, t = 3.31, P = 0.009) and histamine levels(lg histamine:2.13 +/- 0.30 vs 1.45 +/- 0.23, t = 3.34, P = 0.009) of 4% 7 S group at day 19 were all significantly higher, while the serum IFN-gamma content [(35.78 +/- 6.42) pg/ml vs (51.10 +/- 9.67) pg/ml, t = -2.33, P = 0.045] of 4%7 S group was significantly lower.

CONCLUSIONThe WZS miniature swine model orally induced by soybean beta-conglycinin is type I hypersensitivity mediated by IgE, which can be used to predict the potential allergenicity of genetically modified food.

Animals ; Antigens, Plant ; adverse effects ; Disease Models, Animal ; Food Hypersensitivity ; etiology ; Globulins ; adverse effects ; Seed Storage Proteins ; adverse effects ; Soybean Proteins ; adverse effects ; Swine ; Swine, Miniature

Peanut allergy is a significant health problem because of the potential severity of the allergic reaction, the life-long nature of the allergic hypersensitivity, and the ubiquitous use of peanut products. Peanut is one of the most common foods causing serious anaphylactic reactions. Three major peanut allergens, Ara h 1, Ara h 2, and Ara h 3 are 64.5, 17.5, 14 kd glycoproteins belong to the vicillin, conglutin, and glycinin families of seed storage proteins, respectively. We report a case of anaphylaxis induced by peanut with generalized edema, skin rash, dyspnea, hypotension, syncope after accidental peanut ingestion of a 33-year-old man. The diagnosis could be confirmed as peanut-induced anaphylaxis on the basis of prick test, open patch test, MAST, and T cell proliferation assay in addition to history.
Adult ; Allergens ; Anaphylaxis* ; Cell Proliferation ; Diagnosis ; Dyspnea ; Eating ; Edema ; Exanthema ; Glycoproteins ; Humans ; Hypersensitivity ; Hypotension ; Patch Tests ; Peanut Hypersensitivity ; Seed Storage Proteins ; Syncope

Food allergy is an adverse food reaction as a result of immune mechanisms. In a sensitized individual, food allergens activate mast cells and basophils by binding with IgE present on the cell surface, resulting in the release of chemical mediators and various cytokines to cause various clinical symptoms of food allergy. Sensitization to food allergens can occur in the gastrointestinal tract (class 1 food allergy) or as a consequence of cross reactivity to structurally homologous inhalant allergens (class 2 food allergy). The class 1 food allergens are water-soluble glycoproteins with 10-70 kD size that are resistant to heat, acid and enzymes. On the other hand, the class 2 food allergens are highly unstable and degraded by heat or enzymatic digestion. Much progress has been made in identifying and isolating food allergen. Recently cDNAs for many proteins have been isolated and recombinant proteins have been generated. These techniques make it easier to characterize each responsible food allergens. Plant food allergens are classified into families and superfamilies by their structural and functional properties. The most of plant food allergens are the cupin and prolamin superfamilies and the protein families of the plant defense system. The cupin superfamily includes allergenic seed storage proteins of 7s globulin (vicilin) and 11s globulin (legumin). 2s albumin seed storage proteins, the nonspecific lipid transfer proteins, and the cereal alpha-amylase and protease inhibitors belong to the prolamin superfamily. Profilins, heveins, and nonspecific lipid transfer proteins are present in a variety of pollens, nuts, seeds, fruits, and vegetables. These are considered as panallergens, causing a significant degree of IgE-mediated cross-reactivity. Detailed informations about the character of food allergens can be used to develop more sophisticated diagnostic methods and treatment modalities in the near future. Further knowledge of food allergens is also useful to assess the allergenicity of novel protein of genetically mo.
Allergens* ; alpha-Amylases ; Basophils ; Edible Grain ; Classification* ; Cytokines ; Digestion ; DNA, Complementary ; Food Hypersensitivity ; Fruit ; Gastrointestinal Tract ; Glycoproteins ; Hand ; Hot Temperature ; Humans ; Immunoglobulin E ; Mast Cells ; Nuts ; Plants* ; Pollen ; Profilins ; Protease Inhibitors ; Recombinant Proteins ; Seed Storage Proteins ; Vegetables

PURPOSE: The aim of this study was to assess the clinical usefulness and added diagnostic value of specific IgE antibodies to wheat, gluten, and ω-5 gliadin in diagnosing wheat allergy and distinguishing wheat anaphylaxis. METHODS: This study included 196 children who visited Ajou University Hospital for suspicious food allergy. The subjects were divided into 2 groups: the wheat allergy (WA) and non-wheat allergy (non-WA) groups. Patients with wheat allergy were further divided into 2 subgroups according to their symptoms: the wheat allergy with anaphylaxis (WA(Ana)) and wheat allergy without anaphylaxis (WA(Non-Ana)) groups. Serum concentrations of total IgE and specific IgE antibodies to wheat, gluten and ω-5 gliadin were measured. RESULTS: The median values of specific IgE antibodies to wheat, gluten and ω-5 gliadin were significantly higher in the WA group than in the non-WA group, and the positive decision points (95% specificity) were at 3.12, 2.61, and 0.21 kUA/L, respectively. The combination of specific IgE antibodies to wheat and ω-5 gliadin resulted in the highest accuracy of 93.9% in diagnosing wheat allergy. In differentiating the WA(Ana) group from the WA(Non-Ana) group, only specific IgE antibody to ω-5 gliadin showed a significant difference at the optimal cutoff point of 1.56 kUA/L. CONCLUSION: Our results show that the individual levels of specific IgE antibodies to wheat, gluten or ω-5 gliadin may have a considerably high accuracy in diagnosing wheat allergy and that specific IgE antibody to ω-5 gliadin may be particularly useful in predicting wheat anaphylaxis.
Anaphylaxis ; Antibodies ; Child* ; Food Hypersensitivity ; Gliadin* ; Glutens* ; Humans ; Hypersensitivity ; Immunoglobulin E* ; Triticum* ; Wheat Hypersensitivity*

Wheat is a common cause of food allergy. Wheat-induced anaphylaxis (WIA) and wheat-dependent exercise induced anaphylaxis (WDEIA) are severe forms of immunoglobulin E (IgE) mediated allergic reaction to wheat protein. As the diagnosis of WIA or WDEIA is not easy because of the risk of oral challenge, identification of specific IgE of various wheat proteins is helpful for diagnosis. In Korea, there are only a few reports on WIA in adults. We report six cases of WIA diagnosed on the basis of clinical history and specific IgE of wheat proteins or provocation test. For immunologic evaluation of severe wheat allergy including WIA and WDEIA, it is important to measure specific IgE to each component of wheat including gluten and omega-5 gliadin not just measuring wheat-specific IgE.
Adult* ; Anaphylaxis* ; Diagnosis ; Food Hypersensitivity ; Gliadin ; Glutens ; Humans ; Hypersensitivity ; Immunoglobulin E ; Immunoglobulins ; Korea ; Triticum ; Wheat Hypersensitivity

BACKGROUND/AIMS: The role of dietary factors in the pathogenesis of irritable bowel syndrome (IBS) is still unclear. The aim of this study was to compare IgG4 levels to common food antigens between patients with IBS and healthy controls. METHODS: Thirty-two patients diagnosed as IBS according to the Rome III criteria (12 diarrhea subgroup; 20 non-diarrhea subgroup) and 32 sex and age-matched healthy controls participated in the study. Serum IgG4 titers to 90 common foods were measured in each subject. The number of subjects with positivity defined as the cut-off value ≥ 0.7 U/mL was compared. RESULTS: Patients with IBS had significantly higher IgG4 titers to wheat, leek and taro compared to those of controls. Serum IgG4 titers to ginger, cocoa, walnut, white radish, onion, and lettuce in IBS patients tended to be higher than controls. IgG4 titers to wheat, gluten and gliadin in the diarrhea subgroup, and lettuce, leek and taro in the non-diarrhea subgroup tended to be higher compared with controls. The number of subjects with positivity to apple, orange, lettuce, and leek was significantly higher in IBS patients than controls. The number of subjects with positivity to apple, orange, gluten, and gliadin in the diarrhea subgroup, and egg white, pineapple, soybean, lettuce, and leek in the non-diarrhea subgroup was significantly higher compared with controls. CONCLUSIONS: Serum IgG4 antibody levels to some common foods are abnormally elevated in IBS patients. The type of foods with abnormally elevated serum IgG4 titers in the diarrhea subgroup may be different from that in the non-diarrhea subgroup.
Ananas ; Cacao ; Citrus sinensis ; Colocasia ; Diarrhea ; Egg White ; Ginger ; Gliadin ; Glutens ; Humans ; Immunoglobulin G* ; Irritable Bowel Syndrome* ; Juglans ; Lettuce ; Onions ; Raphanus ; Soybeans ; Triticum

BACKGROUND/AIMS: Non-celiac gluten sensitivity has been increasingly recognized as a predisposing factor for irritable bowel syndrome (IBS)-like symptoms in Western populations where celiac disease (CD) is relatively common. In Asia where CD is rare, we wish to determine the prevalence of gluten protein associated serology in IBS patients, which has not been formally studied, and its relation to histological and human leukocyte antigen (HLA) markers. METHODS: We reviewed a consecutive cohort of Asian patients with IBS, who had undergone serologic testing for IgA against deamidated gliadin peptide antibodies (IgA DGP) and IgA anti-endomysium antibodies, and who also had duodenal biopsies during clinical workup. In addition, a subset of Chinese patients with positive serology was further tested for HLA-DQ2 and HLA-DQ8. RESULTS: Of 186 patients, 34 (18%) were positive for IgA DGP; bloating, abdominal pain, belching and diarrhea were the most commonly reported symptoms but diarrhea as the most bothersome symptom was significantly more common in IgA DGP positive patients. Mildly increased intra-epithelial lymphocytes on duodenal biopsy was also more common (29% vs. 9%, P = 0.001). Nine of 21 Chinese patients tested as IgA DGP positive undertook HLA-DQ2/DQ8 testing, with only 2 being positive for HLA-DQ8. All patients with positive IgA DGP reported symptom improvement with gluten withdrawal. CONCLUSIONS: We have described a series of Asian, mainly Chinese, patients with IBS who were tested positive for IgA DGP, and improved on a gluten exclusion diet. We believe this is the first report of non-celiac gluten sensitivity in Asia, a region where CD is uncommon.
Abdominal Pain ; Antibodies* ; Asia ; Asian Continental Ancestry Group* ; Biopsy ; Causality ; Celiac Disease ; Cohort Studies ; Diarrhea ; Diet ; Eructation ; Gliadin* ; Glutens ; Humans ; Immunoglobulin A ; Irritable Bowel Syndrome* ; Leukocytes ; Lymphocytes ; Prevalence* ; Serologic Tests

Wheat is the most widely cultivated grain and an important source of food and dietary protein. Wheat proteins are classified based on extraction in different solvents, which are albumin, globulin, prolamin (gliadin) and glutenin. The term 'gluten' contains approximately equal amounts of gliadin and glutenin and is the major determinant of the properties of wheat flour conferring cohesiveness and viscoelasticity that allows its dough to be processed into many kinds of food. Gluten is known to be responsible for triggering celiac disease and wheat allergy. Wheat allergy is primarily an IgE-mediated response. Clinical manifestations of wheat allergy are similar to those of other food allergies, with symptoms on the skin, gut and respiratory tract. Recent studies have shown that IgE to gliadin can be an indicator for risk of severe immediate reaction-like anaphylaxis and wheat-dependent, exercise-induced anaphylaxis (WDEIA). However, current in vitro test reagents for the diagnosis of wheat allergy mainly contain water-soluble wheat protein and a small amount of gluten, so there are some limitations to diagnose gluten allergy. Furthermore, there is no acceptable method to measure gluten in food products for preparing effective gluten-free diet. To overcome these limitations and to improve quality of life of wheat allergy sufferers, more work is needed. We report a case of a 4-year-old boy with gluten allergy who presented with urticaria after ingestion kneaded wheat flour with a brief review of the literature.
Anaphylaxis ; Celiac Disease ; Edible Grain ; Diet, Gluten-Free ; Dietary Proteins ; Eating ; Flour ; Food Hypersensitivity ; Gliadin ; Glutens ; Hypersensitivity ; Immunoglobulin E ; Indicators and Reagents ; Preschool Child ; Proteins ; Quality of Life ; Respiratory System ; Skin ; Solvents ; Triticum ; Urticaria ; Wheat Hypersensitivity

BACKGROUNDWheat-dependent exercise-induced anaphylaxis (WDEIA) is a complex disease resulting from interaction of environmental and genetic factors. The aim of this study was to investigate the association of three single nucleotide polymorphisms (SNPs) (IL-4-C590T, IL-4RA A1727G and IL-10-A627C) with WDEIA.

METHODSSNP genotyping was conducted among the case subset composing 51 patients with WDEIA and four control subsets by sequencing DNA yielded from polymerase chain reaction (PCR). Statistical analysis of genotype/allele's frequencies between cases and controls were carried out through Fisher's exact test with the software of SPSS16.0.

RESULTSFor IL-4-C590T, there were statistically significant differences of genotype frequencies in case-control 1 (P = 0.03) and case-control 4 (P = 0.001) and statistically significant differences of allele frequencies in three case-control models (case-control 1: OR = 4.27 (95%CI = 1.40 - 13.07), P = 0.009; case-control 3: OR = 1.99 (95%CI = 1.13 - 3.50), P = 0.02; case-control 4: OR = 2.39 (95%CI = 1.49 - 3.84), P = 0.001). All other association studies showed no statistically significant (P > 0.05).

CONCLUSIONSIL-4-C590T may be related to the susceptibility of WDEIA, and the minor allele C might be a potential risk factor accounting for WDEIA. IL-4RA A1727G and IL-10-A627C might not be involved in the occurrence of WDEIA.

Adolescent ; Adult ; Aged ; Anaphylaxis ; genetics ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Exercise ; physiology ; Female ; Gene Frequency ; genetics ; Genotype ; Gliadin ; immunology ; Glutens ; immunology ; Humans ; Immunoglobulin E ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Triticum ; immunology ; Young Adult