Search for atoxic cereals: a single blind, cross-over study on the safety of a single dose of Triticum monococcum, in patients with celiac disease

Abstract

BackgroundCereals of baking quality with absent or reduced toxicity are actively sought as alternative therapy to a gluten-free diet (GFD) for patients with coeliac disease (CD). Triticum monococcum, an ancient wheat, is a potential candidate having no toxicity in in-vitro and ex-vivo studies. The aim of our study was to investigate on the safety of administration of a single dose of gluten of Tm in patients with CD on GFD.MethodsWe performed a single blind, cross-over study involving 12 CD patients who had been on a GFD for at least 12 months, challenged on day 0, 14 and 28 with a single fixed dose of 2.5 grams of the following (random order): Tm, rice (as reference atoxic protein) and Amygluten (as reference toxic protein) dispersed in a gluten-free pudding. The primary end-point of the study was the change in intestinal permeability, as assessed by changes in the urinary lactulose/rhamnose ratio (L/R ratio) measured by High Pressure Liquid Chromatography. We also assessed the occurrence of adverse gastrointestinal events, graded for intensity and duration according to the WHO scale. Variables were expressed as mean ± SD; paired t-test and χ2 test were used as appropriate.ResultsThe urinary L/R ratio did not change significantly upon challenge with the 3 cereals, and was 0.055 ± 0.026 for Tm Vs 0.058 ± 0.035 for rice (p = 0.6736) and Vs 0.063 ± 0.054 with Amygluten (p = 0.6071). Adverse gastrointestinal events were 8 for Tm, Vs 11 for rice (p = 0.6321) and Vs 31 for Amygluten p = 0.0016), and, in all cases events were graded as “mild” or “moderate” with TM and rice, and as “severe” or “disabling” in 4 cases during Amygluten.ConclusionsNo definite conclusion can be drawn on the safety of Tm, based on no change in urinary L/R because even Amygluten, a toxic wheat protein, did not cause a significant change in urinary L/R indicating low sensitivity of this methodology in studies on acute toxicity. Tm was, however, well tolerated by all patients providing the rationale for further investigation on the safety of this cereal for CD patients.Trial registrationEudraCT-AIFA n2008-000697-20

In Celiac Disease Patients the In Vivo Challenge with the Diploid Triticum monococcum Elicits a Reduced Immune Response Compared to Hexaploid Wheat

“…[ 21 ] Nevertheless, the large individual variability and the residual immunological activity confirm that TM is a cereal not suitable for the diet of CD patients, as reported in previous studies. [ 22,23 ] Actually, our in vivo results are in agreement with Zanini and colleagues, who have conducted the unique two in vivo studies to address the effects of T. monococcum ( Monlis cultivar) in CD subjects, concluding that TM is toxic for CD patients. [ 22,23 ] In addition, our findings support the validity of the SGC as a tool to monitor in CD patients the immune responses to naturally or in vitro detoxified cereals.…”

“…[ 22,23 ] Actually, our in vivo results are in agreement with Zanini and colleagues, who have conducted the unique two in vivo studies to address the effects of T. monococcum ( Monlis cultivar) in CD subjects, concluding that TM is toxic for CD patients. [ 22,23 ] In addition, our findings support the validity of the SGC as a tool to monitor in CD patients the immune responses to naturally or in vitro detoxified cereals. [ 40,41 ]…”

“…Its capability to trigger immunity in CD patients has been investigated by several in vitro, ex vivo and recently also by two in vivo studies, that overall showed conflicting results. [ 15–23 ] TM lacks the D genome that encodes for α‐gliadin proteins containing the 33‐mer peptide, highly immunostimulatory in CD patients. [ 12–14 ] Moreover, recent studies reported that some TM cultivars have a ω‐gliadin peptide able to prevent the “toxic” effects induced by gluten proteins in in vitro cellular and mucosal models.…”

Wheat Seed Proteins: Factors Influencing Their Content, Composition, and Technological Properties, and Strategies to Reduce Adverse Reactions

“…Assays of these genotypes with intestinal T-cell clones derived from the biopsy samples of coeliac patients showed almost complete suppression of disease-related T-cell epitopes (Gil-Humanes et al, 2010). More recently, the same group used the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing technology to target conserved regions adjacent to the coding sequence of the 33-mer peptide in the α2-gliadin genes α/β-gliadins (Gli-A2) and/or γ -and No negative effects on enterocyte height (Frisoni et al, 1995;Lafiandra, Colaprico, Kasarda, & Porceddu, 1987) α/β-gliadins (Gli-A2) and (Autran, 1975;Branlard, Dardevet, Amiour, & Igrejas, 2003; β-, γ -, and ω-gliadins Activated CD T-cell response (Gianfrani et al, 2012) Urinary lactulose/ rhamnose ratio (L/R ratio measured by high-pressure liquid chromatography Well tolerated by all patients (Zanini et al, 2013) α/β-, γ -, or (Ingversen, Koie, & Doll, 1973;Sestak et al, 2015Sestak et al, , 2016 lys3b ( ELISA and mass spectrometry Grain has application in the preparation of food and beverages for coeliacs (Tanner, Blundell, Colgrave, & Howitt, 2016) lys1 (5HL) and lys3a (5HS) DH lines (Risø 1508 x Hiproly)…”

Gene editing towards hypoimmunogenic gluten proteins in wheat

“…Diploid einkorn wheat (T. monococcum, A genome) has good nutritional characteristics and bread making quality. Food products made from the variety 'Monlis' have been shown to be tolerated by CD patients, revealing similar absence of CD toxicity-related symptoms as found for rice (Zanini et al, 2013). New combinations of gluten genes can be made by crossing and selection at the diploid level.…”