Tissue transglutaminase, the celiac disease autoantigen, preferentially generates antigenic neoepitopes in γ and γ but not in α gliadins

Rothia mucilaginosa, a natural microbial inhabitant of the oral cavity, cleaves gluten (gliadin) proteins at regions that are resistant to degradation by mammalian enzymes. The aim of this study was to investigate to what extent the R. mucilaginosa cell-associated enzymes abolish gliadin immunogenic properties. Degradation of total gliadins and highly immunogenic gliadin 33-mer or 26-mer peptides was monitored by SDS-PAGE and RP-HPLC, and fragments were sequenced by liquid chromatography and electrospray ionization tandem mass spectrometer (LC-ESI-MS/MS). Peptide deamidation by tissue transglutaminase (TG2), a critical step in rendering the fragments more immunogenic, was assessed by TG2-mediated cross-linking to monodansyl cadaverine (MDC), and by a +1-Da mass difference by LC-ESI-MS. Survival of potential immunogenic gliadin epitopes was determined by use of the R5 antibody-based ELISA. R. mucilaginosa-associated enzymes cleaved gliadins, 33-mer and 26-mer peptides into smaller fragments. TG2-mediated cross-linking showed a perfect inverse relationship with intact 33-mer and 26-mer peptide levels, and major degradation fragments showed a slow rate of MDC cross-linking of 6.18 ± 2.20 AU/min compared with 97.75 ± 10.72 and 84.17 ± 3.25 AU/min for the intact 33-mer and 26-mer, respectively, which was confirmed by reduced TG2-mediated deamidation of the fragments in mass spectrometry. Incubation of gliadins with Rothia cells reduced R5 antibody binding by 20, 82, and 97% after 30 min, 2 h, and 5 h, respectively, which was paralleled by reduced reactivity of enzyme-treated 33-mer and 26-mer peptides in the R5 competitive ELISA. Our broad complementary approach to validate gluten degrading activities qualifies R. mucilaginosa-associated enzymes as promising tools to neutralize T cell immunogenic properties for treatment of celiac disease.

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INCREASED TISSUE TRANSGLUTAMINASE LEVELS ARE ASSOCIATED WITH INCREASED EPILEPTIFORM ACTIVITY IN ELECTROENCEPHALOGRAPHY AMONG PATIENTS WITH CELIAC DISEASE

Arquivos de Gastroenterologia ◽

10.1590/s0004-28032015000400005 ◽

2015 ◽

Vol 52 (4) ◽

pp. 272-277 ◽

Cited By ~ 3

Author(s):

Sedat IŞIKAY ◽

Şamil HIZLI ◽

Serkan ÇOŞKUN ◽

Kutluhan YILMAZ

Keyword(s):

Celiac Disease ◽

Tissue Transglutaminase ◽

Pearson Correlation ◽

Epileptiform Activity ◽

Control Group ◽

Healthy Children ◽

Newly Diagnosed ◽

Laboratory Findings ◽

Significant Difference ◽

Medical Histories

Background - Celiac disease is an autoimmune systemic disorder in genetically predisposed individuals precipitated by gluten ingestion. Objective - In this study, we aimed to determine asymptomatic spike-and-wave findings on electroencephalography in children with celiac disease. Methods - A total of 175 children with the diagnosis of celiac disease (study group) and 99 age- and sex-matched healthy children as controls (control group) were included in the study. In order to determine the effects of gluten free diet on laboratory and electroencephalography findings, the celiac group is further subdivided into two as newly-diagnosed and formerly-diagnosed patients. Medical histories of all children and laboratory findings were all recorded and neurologic statuses were evaluated. All patients underwent a sleep and awake electroencephalography. Results - Among 175 celiac disease patients included in the study, 43 were newly diagnosed while 132 were formerly-diagnosed patients. In electroencephalography evaluation of patients the epileptiform activity was determined in 4 (9.3%) of newly diagnosed and in 2 (1.5%) of formerly diagnosed patients; on the other hand the epileptiform activity was present in only 1 (1.0%) of control cases. There was a statistically significant difference between groups in regards to the presence of epileptiform activity in electroencephalography. Pearson correlation analysis revealed that epileptiform activity in both sleep and awake electroencephalography were positively correlated with tissue transglutaminase levels (P=0.014 and P=0.019, respectively). Conclusion - We have determined an increased epileptiform activity frequency among newly-diagnosed celiac disease patients compared with formerly-diagnosed celiac disease patients and control cases. Moreover the tissue transglutaminase levels were also correlated with the presence of epileptiform activity in electroencephalography. Among newly diagnosed celiac disease patients, clinicians should be aware of this association and be alert about any neurological symptoms.

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Relationships between Clinical Presentation, Serology, Histology, and Duodenal Deposits of Tissue Transglutaminase Antibodies in Pediatric Celiac Disease

Digestive Diseases ◽

10.1159/000490377 ◽

2018 ◽

Vol 36 (5) ◽

pp. 369-376 ◽

Cited By ~ 1

Author(s):

Nurit Loberman-Nachum ◽

Michael Schvimer ◽

Camila Avivi ◽

Iris Barshack ◽

Avishay Lahad ◽

...

Keyword(s):

Celiac Disease ◽

Immunohistochemical Staining ◽

Clinical Presentation ◽

Tissue Transglutaminase ◽

Mucosal Damage ◽

High Serum ◽

Tissue Transglutaminase Antibodies ◽

Antibody Levels ◽

Age Range ◽

Multiple Symptoms

Background: The clinical, histological, and serological spectrum of celiac disease (CD) vary widely. We aimed to examine relationships between symptoms, serum anti-tissue transglutaminase antibodies (tTG) levels, mucosal damage, and mucosal anti-tTG deposits in pediatric CD. Methods: A retrospective single-center, cohort study of children referred for endoscopy with suspected CD during 2011–2014. We retrieved the clinical data, blindly reviewed duodenal biopsies, and performed immunohistochemical staining for anti-tTG deposits. Patients were classified as monosymptomatic or polysymptomatic. Mucosal anti-tTG deposits were classified according to the location of deposits, dominant intensity, maximal intensity, and percentage of stained area. Results: Of 252 patients with confirmed CD, complete data were available for 100: 37 males in the age range 1.3–16.7 with median 4.0 years. Monosymptomatic patients (n = 54) presented at an older age than polysymptomatic patients (1.3–15.5, median 8.1 vs. 1.3–16.7, median 6.3 years, p = 0.026). Marsh 2–3c was more prevalent in polysymptomatic patients (93 vs. 78%, p = 0.028). The intensity of mucosal anti-tTG deposits correlated with serum anti-tTG levels but not with the clinical presentation. Conclusions: Multiple symptoms and high serum anti-tTG antibody levels correlated with mucosal damage in children with CD. The role of immunohistochemical staining for intestinal anti-tTG mucosal deposits in the diagnosis of borderline CD is not yet established.

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Screening Tests Using Serum Tissue Transglutaminase IgA May Facilitate the Identification of Undiagnosed Celiac Disease among Japanese Population

International Journal of Medical Sciences ◽

10.7150/ijms.8854 ◽

2014 ◽

Vol 11 (8) ◽

pp. 819-823 ◽

Cited By ~ 8

Author(s):

Hideyuki Nakazawa ◽

Hideki Makishima ◽

Toshiro Ito ◽

Hiroyoshi Ota ◽

Kayoko Momose ◽

...

Keyword(s):

Celiac Disease ◽

Japanese Population ◽

Tissue Transglutaminase ◽

Screening Tests

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Prevalence of Celiac Disease in Collagenous and Lymphocytic Colitis

Canadian Journal of Gastroenterology ◽

10.1155/2000/847807 ◽

2000 ◽

Vol 14 (11) ◽

pp. 919-921 ◽

Cited By ~ 31

Author(s):

Helen Rachel Gillett ◽

Hugh James Freeman

Keyword(s):

Celiac Disease ◽

Small Bowel ◽

Tissue Transglutaminase ◽

Immunoglobulin A ◽

Collagenous Colitis ◽

Lymphocytic Colitis ◽

Small Bowel Biopsy ◽

Major Antigen ◽

Immunofluorescent Technique ◽

Endomysium Antibody

Both collagenous and lymphocytic colitis have been described in patients with celiac disease, suggesting an association between the conditions. Over the past few years, the availability, sensitivity and specificity of serological markers for celiac disease have improved - the most recent advancement being the description of tissue transglutaminase as the major antigen for endomysium antibody. A quantitative ELISA was used to measure titres of immunoglobulin A (IgA) antibody to tissue transglutaminase (tTG) along with an immunofluorescent technique for IgA endomysium antibody (EmA) in 15 patients with lymphocytic colitis and eight with collagenous colitis to determine whether celiac disease latency could be detected. One patient with lymphocytic colitis demonstrated both elevated titres of tTG antibody and positive EmA, and small bowel biopsy confirmed celiac disease. One patient with collagenous colitis had a slightly elevated titre of tTG antibody with a negative EmA, and results of a small bowel biopsy were normal. Three other patients with lymphocytic colitis were already treated for previously diagnosed celiac disease. The prevalence of celiac disease occurring in lymphocytic colitis was found to be 27%, but no cases of celiac disease in association with collagenous colitis were found.

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