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

Nakamura, Ryosuke; Nakamura, Rika; Sakai, Shinobu; Adachi, Reiko; Hachisuka, Akiko; Urisu, Atsuo; Fukutomi, Yuma; Teshima, Reiko

doi:10.1016/j.jaci.2013.07.017

Cited by 25 publications

(19 citation statements)

References 7 publications

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“…The higher allergenic potency of GP19S and 0.5 hr acid-HWP compared with gluten seems to stem from the deamidation and solubility increase with similar molecular weight (supplementary Fig. S1, Nakamura et al, 2013b). Predictably, the allergenic potency of GP19S was higher than that of gluten in the EC sensitized guinea pig model.…”

Section: Discussionmentioning

confidence: 91%

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

et al. 2015

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-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.

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Section: Discussionmentioning

confidence: 91%

“…The allergenicity of HWP is strongly influenced by the deamidation and molecular weight of the protein (Chinuki et al, 2013;Gourbeyre et al, 2014;Nakamura et al, 2013b). In addition, acid hydrolysis increases an aqueous solubility of the gluten.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2015

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“…[7][8][9][10] Briefly, the cells 4 cells/50 µL/well) were sensitized with 10 ng/mL anti-OVA IgE Abs and plated on a clear-bottom white 96-well plate (ViewPlate; Perkin Elmer, Inc.). After overnight incubation, the cells were washed three times with PBS and then stimulated with OVA diluted in MEM containing 10% FCS (50 µL/well) at 37°C under an atmosphere of 5.0% CO 2 for 3 h. After stimulation, 50 µL of luciferase substrate solution containing cell lysis reagent (ONE-Glo; Promega Corporation, Madison, WI, U.S.A.) was added to the cells, and chemiluminescence was measured using an EnVision multilabel plate reader (PerkinElmer, Inc.).…”

Section: Reagentsmentioning

confidence: 99%

“…To overcome this blocking effect, excessive amounts of antigens are immobilized in the solid phase of immunochemical allergy tests, such as the radioallergosorbent test and fluoroenzyme immunoassay ImmunoCAP ® (CAP test). 4,5) However, immobilization may affect the physicochemical nature of the antigens, resulting in discrepancies of the allergenicity of the molecule between in vivo responses and in vitro tests.We developed a new allergy test, called the IgE crosslinking-induced luciferase expression (EXiLE) test, [6][7][8][9][10] which is dependent on the activation of the humanized rat mast cell line RS-ATL8 sensitized with serum IgE of allergic patients. The EXiLE test detects the ability of IgE to crosslink multivalent antigens in the liquid phase, resulting in a more reliable diagnosis of egg white food allergy than the conventional CAP test.…”

mentioning

confidence: 99%

“…We developed a new allergy test, called the IgE crosslinking-induced luciferase expression (EXiLE) test, [6][7][8][9][10] which is dependent on the activation of the humanized rat mast cell line RS-ATL8 sensitized with serum IgE of allergic patients. The EXiLE test detects the ability of IgE to crosslink multivalent antigens in the liquid phase, resulting in a more reliable diagnosis of egg white food allergy than the conventional CAP test.…”

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Different Results of IgE Binding- and Crosslinking-Based Allergy Tests Caused by Allergen Immobilization

Okamoto‐Uchida¹,

Nakamura²,

Y³

et al. 2016

Biological & Pharmaceutical Bulletin

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The physicochemical nature of allergen molecules differ from the liquid phase to the solid phase. However, conventional allergy tests are based on the detection of immunoglobulin (Ig)E binding to immobilized allergens. We recently developed an in vitro allergy testing method using a luciferase-reporting humanized rat mast cell line to detect IgE crosslinking-induced luciferase expression (EXiLE test). The aim of the present study was to evaluate the effects of antigen immobilization on the results of different in vitro allergy tests using two anti-ovalbumin (OVA) antibodies (Abs), E-C1 and E-G5, with different properties in the OVAinduced allergic reaction. Both Abs showed clear binding to OVA with an enzyme-linked immunosorbent assay and by BIAcore analysis. However, only E-C1 potentiated EXiLE response for the liquid-phase OVA. On the other hand, OVA immobilized on solid-phase induced EXiLE responses in both E-C1 Ab-and E-G5 Ab-sensitized mast cells. Western blotting of OVA indicated that E-C1 Ab binds both to OVA monomers and dimers, unlike E-G5 Ab, which probably binds only to the OVA dimer. These results suggest that antigen immobilization enhanced IgE crosslinking ability through multimerization of allergen molecules in the solid phase, resulting in an increase in false positives in IgE binding-based conventional in vitro allergy tests. These findings shed light on the physicochemical nature of antigens as an important factor for the development and evaluation of in vitro allergy tests and suggest that mast cell activation-based allergy testing with liquid-phase allergens is a promising strategy to evaluate the physiological interactions of IgE and allergens.Key words immunoglobulin E (IgE); allergen; immobilization; in vitro allergy test; luciferase assay; IgE crosslinking-induced luciferase expression (EXiLE) test Immunoglobulin (Ig) E is the primary factor for mast cell activation and type I allergic reactions, such as histamine release and production of inflammatory factors. Although immunochemical allergen-specific IgE tests based on the binding of serum IgE to an allergen are used worldwide, the results cannot always be translated into a clear diagnosis, especially in the case of food allergies.1,2) The existence of allergen-specific IgE indicates that the individual has been previously sensitized to the allergen. However, IgE is not always capable of crosslinking high-affinity IgE receptors (FcεRI) with the soluble allergen because multiple epitopes of an allergen molecule are required.3) Additionally, there is a greater abundance of antigen-specific IgGs in sera than IgE, which may interfere with the binding of IgE to the antigen. To overcome this blocking effect, excessive amounts of antigens are immobilized in the solid phase of immunochemical allergy tests, such as the radioallergosorbent test and fluoroenzyme immunoassay ImmunoCAP ® (CAP test). 4,5) However, immobilization may affect the physicochemical nature of the antigens, resulting in discrepancies of the allergenicity of the molecule betw...

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Total dose defines the incidence of percutaneous IgE/IgG1 mediated immediate‐type hypersensitivity caused by papain

Yokozeki

Tobisawa

Ogasawara

et al. 2020

J of Applied Toxicology

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Assessment of human health risk requires an understanding of antigen dose metrics associated with toxicity. Whereas assessment of the human health risk for delayed‐type hypersensitivity is understood, the metrics remain unclear for percutaneous immediate‐type hypersensitivity (ITH) mediated by IgE/IgG1. In this work, we aimed to investigate the dose metric for percutaneous ITH mediated by IgE/IgG1 responses. Papain, which causes ITH via percutaneous sensitization in humans, was used to sensitize guinea pigs and mice. The total dose per animal or dose per unit area was adjusted to understand the drivers of sensitization. Passive cutaneous anaphylaxis (PCA) and enzyme‐linked immunosorbent assay (ELISA) for papain‐specific IgG1 enabled quantification of the response in guinea pigs. In mice, the number of antigen‐bearing B cells in the draining lymph nodes (DLN) was calculated using flow cytometry papain‐specific IgG1 and IgE levels were quantified by ELISA. PCA positive test rates and the amounts of antigen‐specific antibody corresponded with total dose per animal, not dose per unit area. Furthermore, the number of B cells taking up antigen within DLN also correlated with total dose. These findings indicate that the total antigen dose is the important metric for percutaneous IgE/IgG1‐mediated ITH.

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

Cited by 25 publications

References 7 publications

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

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

Different Results of IgE Binding- and Crosslinking-Based Allergy Tests Caused by Allergen Immobilization

Total dose defines the incidence of percutaneous IgE/IgG1 mediated immediate‐type hypersensitivity caused by papain

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