Use of ancient wheat crops for the diet of non-celiac gluten sensitive patients
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1 Use of ancient wheat crops for the diet of non-celiac gluten sensitive patients Giuseppe Mazzarella Institute of Food Sciences-CNR - Avellino 9th PROBIOTICS, PREBIOTICS & NEW FOODS, NUTRACEUTICALS AND BOTANICALS for Nutrition & Human and Microbiota Health Roma, 10/12 Settembre 2017
2 Proposed New Classification of Gluten Related Disorders Biomarkers Gluten Related Disorders Pathogenesis YES Autoimmune Allergic NO Not Autoimmune Not allergic (Innate immunity?) Celiac Disease Adaptive and + Innate Immunity Gluten Ataxia Dermatitis herpetiformis YES Wheat allergy Gluten sensitivity Symptomitic Silent Potential Respiratory Allergy Food Allegy WDEIA Contact Urticaria
3 The clinical manifestations of Non-Celiac Gluten Sensitivity (NCGS). Frequency Intestinal Extra-intestinal Very Common Bloating Lack of wellbeing Abdominal pain Tiredness Common Diarrhea Headache Epigastric pain Anxiety Nausea Foggy mind Aerophagia Numbness GER Joint/muscle pain Aphtous stomatitis Skin rash/dermatitis Alternating bowel habits Weight loss Constipation Anemia Loss of balance Undetermined Hematochezia Depression Anal fissures Rhinitis/asthma Weight increase Interstitial cystitis Ingrown hairs Oligo or polimenorrhea Sensory symptoms Disturbed sleep pattern Hallucinations Mood swings Autism Schizophrenia
4 DIAGNOSIS CD: serology ( anti-tg2) and histology ( villous atrophy) WA: wheat specific IgE antibodies NCGS: absence of specific diagnostic markers To date, is mainly a diagnosis of exclusion
5 Pathogenesis of Celiac Disease (CD)
6 Pathogenesis NCGS innate response? Sapone A et al. Int Arch Allergy Immunol Sapone A et al. BMC 2011 Maria I. Vazquez-Roque et al. Gastroenterology. 2013
7 A B Duodenal IHC CD3+ IELs A small increased of CD3+ intraepithelial lymphocytes (IELs) could be detected in some NCGS patients, compared to controls, C 40 X A= Gluten Sensitive B= Controls C= CD Active Sapone A. et alt, Int. Arch. of Allergy and Immunology 2009
8 The genealogy of cultivated members of the Triticum family Research is intense to find wheat varieties with absent or low toxicity to be implemented in new strategies for treatment and prevention of CD. Among candidates, there are ancient, diploid wheat species for their low capability to activate intestinal T cell responses in CD patients
9 Values reported in the literature within and among wheat types for celiac reactivity Molberg and others 2005; Pizzuti and others 2006; Vincentini and others 2007, 2009; van den Broeck and others 2010a, 2010b
10 Absence of immunotoxic 33-mer peptide in Triticum Monococcum Einkorn
11 Treatment with gliadin peptides from two monococcum accessions induces γifn production from all intestinal gliadin-specific T-cell lines. PT-Sag TG PT-Monlis TG 3500 Dose-response curves PT-ID-331 TG pg/ml IFN-g ug/ml PT
12 Challenge with gliadin peptides from two monococcum accessions induced increase of IEL and LP CD25 cells in treated CD biopsies. IMMUNOHISTOCHEMISTRY Lamina propria CD25+ cells IEL CD3+ cells CD25+ cells/mm 2 lamina propria ** * * CD3+ cells/mm epiteluim ** ** * 0 T. aestivum wheat giadin T. monococcum monlis gliadin T. monococcum ID-331 gliadin 0 T. aestivum wheat giadin T. monococcum monlis gliadin T. monococcum ID-331 gliadin
13 Gliadins from monococcum wheat Monlis but not those from monococcum wheat ID331 induced IL-15 expression in intestinal mucosa from treated CD patients. Medium PT-ID331 PT-Monlis Medium P TG PT- monlis PT- ID331
14 SUMMARY Our data show that the monococcum lines Monlis and ID331 activate the CD T cell response and suggest that these lines are toxic for celiac patients. However, ID331 is likely to be less effective in inducing CD because of its inability to activate the innate immune pathways. Gianfrani C et al. Am J Clin Nutr 2012;96:
15 Gliadin proteins RP-HPLC analysis PC GD-BBM Collected fraction Pepsin (2h) Chymotrypsin (4h) Pepsin (30 min) Trypsin, Chymotrypsin, Elastase, Carboxypeptidase (1h) Brush Border Membrane enzymes (BBM) (2h) Chymotrypsin (16h) nlc-ms/ms Data base search GD-BBM Pepsin (30 min) Trypsin, Chymotrypsin, Elastase, Carboxypeptidase (1h) Brush Border Membrane enzymes (BBM) (2h) Deamindation by ttgase (2h) nlc-ms/ms T-cell and biopsy assay Gliadin identification Data base search Immunogenity evaluation Immunassay Identification of resistant peptides Proteomics
16 Marked reduction of specific T-cell response to monococcum gliadin after the GD-BBM digestion 1200 Pt#CD Interferon-g (pg/ml) PC GD-BBM I331-PC I331 GI-BBM Monlis PC Monlis GI-BBM Sag Marked reduction of T cell activation response to monococcum gliadin after the GD-BBM digestion CD3+CD25+ activated T cells * p<0,001 * p<0,005 * p<0,005 A CD 25+ cells\mm² lamina propria * p<0,005 * p<0,003 * p<0,05 0 Sag-PC Sag GD-BBM ID331-PC ID331 GD-BBM 0 Sag-PC Sag GD-BBM ID331-PC ID331 GD-BBM
17 ID331 MS-based analysis showed that several Triticum monococcum peptides, including Monlis known T-cell epitopes, were degraded during the gastrointestinal treatment, whereas many of Triticum aestivum gliadin survived the gastrointestinal digestion. Sagittario w a/b g
18 SUMMARY Therefore our data confirm that monocoque grain is not a safe grain for celiac disease, but could be a cereal candidate to represent a major food alternative for patients with NCGS and / or for the prevention of celiac disease in subjects at risk for familiarity. Mazzarella G et al. Mol Nutr Food Res Sep;59(9):
19 Non-gluten α -Amylase/Trypsin Inhibitor (ATI) family are strong activators of innate immunity. Dose response of IL-8 release by monocytederived DCs stimulated with purified ATI. LPS served as positive control Junker Y et al J Exp Med 2012;209(13):
20 ATI bioactivity determination in ancient and modern cereals Victor F. Zevallos et al. Gastroenterology 2017;152:
21 Triticum Monococcum as a healthy food The properties of MonLis einkorn compared with a variety of common wheat: higher protein content; higher content of ash and trace elements (iron, zinc, magnesium, phosphorus, potassium etc.); low presence of saturated fatty acids; lower starch content and better digestibility; higher presence of antioxidants and yellow pigmentation (carotenoids, including beta-carotene, the precursor to vitamin A); higher content of tocols (vitamin E). These facts are relevant to the functional activities of cells and are efficient antioxidants Therefore, these data suggest their potentiality as functional foods
22 Thanks for your attention Acknowledgments Carmen Gianfrani Gianfranco Mamone Vera Rotondi Aufiero ISA-CNR, Avellino Salvatore Auricchio Riccardo Troncone Dept Pediatrics, and ELFID Univ Federico II, Naples Alessio Fasano Massachusetts General Hospital Harvard Medical School Nicola Giardullo Gaetano Iaquinto U.O. Gastroenterolgy, Hosp Moscati, Avellino
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