The Preferred Substrates for Transglutaminase 2 in a Complex Wheat Gluten Digest Are Peptide Fragments Harboring Celiac Disease T-Cell Epitopes

Dørum, Siri; Arntzen, Magnus Ø.; Qiao, Shuo‐Wang; Holm, Anders; Koehler, Christian; Thiede, Bernd; Sollid, Ludvig M.; Fleckenstein, Burkhard

doi:10.1371/journal.pone.0014056

Cited by 77 publications

(78 citation statements)

References 35 publications

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“…After SDS-PAGE, the Coomassie-stained H chain bands were excised and in-gel digested with trypsin or chymotrypsin overnight at 37˚C. The proteolytic fragments were extracted from the gel, and biotinylated peptides were isolated with streptavidin-coated Dynabeads as previously described (20). The cross-linked fragments were detected and identified by MALDI-TOF and MALDI-TOF/TOF mass spectrometry using an Ultraflex II instrument (Bruker Daltonics).…”

Section: Identification Of Target Lysine Residuesmentioning

confidence: 99%

Igs as Substrates for Transglutaminase 2: Implications for Autoantibody Production in Celiac Disease

Iversen

Pré

Niro

et al. 2015

The Journal of Immunology

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Autoantibodies specific for the enzyme transglutaminase 2 (TG2) are a hallmark of the gluten-sensitive enteropathy celiac disease. Production of the Abs is strictly dependent on exposure to dietary gluten proteins, thus raising the question how a foreign Ag (gluten) can induce an autoimmune response. It has been suggested that TG2-reactive B cells are activated by gluten-reactive T cells following receptor-mediated uptake of TG2–gluten complexes. In this study, we propose a revised model that is based on the ability of the BCR to serve as a substrate to TG2 and become cross-linked to gluten-derived peptides. We show that TG2-specific IgD molecules are preferred in the reaction and that binding of TG2 via a common epitope targeted by cells using the IgH variable gene segment (IGHV)5–51 results in more efficient cross-linking. Based on these findings we hypothesize that IgD-expressing B cells using IGHV5–51 are preferentially activated, and we suggest that this property can explain the previously reported low number of somatic mutations as well as the overrepresentation of IGHV5–51 among TG2-specific plasma cells in the celiac lesion. The model also couples gluten peptide uptake by TG2-reactive B cells directly to peptide deamidation, which is necessary for the activation of gluten-reactive T cells. It thereby provides a link between gluten deamidation, T cell activation, and the production of TG2-specific Abs. These are all key events in the development of celiac disease, and by connecting them the model may explain why the same enzyme that catalyzes gluten deamidation is also an autoantigen, something that is hardly coincidental.

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Section: Identification Of Target Lysine Residuesmentioning

confidence: 99%

Igs as Substrates for Transglutaminase 2: Implications for Autoantibody Production in Celiac Disease

Iversen

Pré

Niro

et al. 2015

The Journal of Immunology

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“…Only peptides with proteolytic stability are expected to survive this digestion. Second, the digest was treated with TG2, which has been demonstrated to be an important step in the generation of gluten T cell epitopes (32). Finally, the TG2-treated peptides were allowed to bind to HLA-DQ molecules in the presence of HLA-DM, which facilitates peptide exchange and loading.…”

Section: Discussionmentioning

confidence: 99%

HLA-DQ Molecules as Affinity Matrix for Identification of Gluten T Cell Epitopes

Dørum

Bodd

Fallang

et al. 2014

The Journal of Immunology

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Even though MHC class II is a dominant susceptibility factor for many diseases, culprit T cell epitopes presented by disease-associated MHC molecules remain largely elusive. T cells of celiac disease lesions recognize cereal gluten epitopes presented by the disease-associated HLA molecules DQ2.5, DQ2.2, or DQ8. Employing celiac disease and complex gluten Ag digests as a model, we tested the feasibility of using DQ2.5 and DQ2.2 as an affinity matrix for identification of disease-relevant T cell epitopes. Known gluten T cell epitope peptides were enriched by DQ2.5, whereas a different set of peptides was enriched by DQ2.2. Of 86 DQ2.2-enriched peptides, four core sequences dominated. One of these core sequences is a previously known epitope and two others are novel epitopes. The study provides insight into the selection of gluten epitopes by DQ2.2. Furthermore, the approach presented is relevant for epitope identification in other MHC class II–associated disorders.

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“…17,18 However, there are other gluten peptides also in the W-gliadin group which have immunomodulatory activities. [18][19][20][21][22] Although a gluten-free diet is the cure for gluten sensitivity, life on a gluten-free diet can be difficult due to several reasons-expense, contamination of gluten-free products with gluten, products with gluten-derived ingredients, availability, inadequate food labeling, and insufficient knowledge about gluten. Complications of untreated CDs or gluten intolerance can be severe.…”

Section: Staphylococcus Epidermismentioning

confidence: 99%

Rapid isolation of gluten-digesting bacteria from human stool and saliva by using gliadin-containing plates

Berger¹,

Sarantopoulos²,

Ongchangco³

et al. 2014

Exp Biol Med (Maywood)

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The number of individuals with gluten intolerance has increased dramatically over the last years. To date, the only therapy for gluten intolerance is the complete avoidance of dietary gluten. To sustain a strictly gluten-free diet, however, is very challenging. Therefore, there is need for a non-dietary therapy. Any such treatment must appreciate that the immunogenic part of gluten are gliadin peptides which are poorly degraded by the enzymes of the gastrointestinal tract. Probiotic therapy and oral enzyme therapy containing gluten-degrading bacteria (GDB) and their gliadin-digesting enzymes are possible new approaches for the treatment of gluten intolerance, however effectively isolating GDB for these treatments is problematic. The goal of this study was to develop an easy technique to isolate GDB rapidly and efficiently with the hope it might lead to newer ways of developing either probiotics or traditional medicines to treat gluten intolerance. Several researchers have already isolated successfully GDB by using gluten minimal or limited agar plates. Although these plates can be used to isolate bacteria which can tolerate gluten, further assays are needed to investigate if the same bacteria can also digest gluten. The agar plates we developed can detect bacteria which cannot only tolerate gluten but are able to digest it as well. Therefore, we were able to combine two steps into one step. Using such technologies, we were able to isolate five GDB from saliva and stool, and identified three bacterial reference strains with gluten-degrading activity. The technique we developed to isolate bacteria with gluten-degrading activity is fast, effective, and easy to use. The GDB isolated by our technology could have potential as part of a probiotic or enzymatic therapy for people with gluten intolerance.

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The Preferred Substrates for Transglutaminase 2 in a Complex Wheat Gluten Digest Are Peptide Fragments Harboring Celiac Disease T-Cell Epitopes

Cited by 77 publications

References 35 publications

Igs as Substrates for Transglutaminase 2: Implications for Autoantibody Production in Celiac Disease

Igs as Substrates for Transglutaminase 2: Implications for Autoantibody Production in Celiac Disease

HLA-DQ Molecules as Affinity Matrix for Identification of Gluten T Cell Epitopes

Rapid isolation of gluten-digesting bacteria from human stool and saliva by using gliadin-containing plates

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