Wheat Gliadin Research Articles

Wheat gliadins modified by deamidation are more efficient than native gliadins in inducing a Th2 response in Balb/c mice experimentally sensitized to wheat allergens

Wheat gluten proteins such as gliadins constitute major food allergens. Gluten can be modified industrially by deamidation which increases its solubility and enhances its use as a food ingredient. Sensitization to deamidated gluten has been reported to cause severe allergic reactions with anaphylaxis. The aim of this study was therefore to compare the sensitization and elicitation potentials of native (NG) and deamidated (DG) gliadins. The reactivity pattern of mice IgE was also compared with that of DG-allergic patients. The ability of DG to sensitize Balb/c mice using intra-peritoneal administration with aluminium hydroxide as an adjuvant, and to elicit an allergic response after a challenge, was tested in comparison with NG. Mice sensitized with DG secreted higher levels of total IgE, IL-4, gliadin-specific IgE and IgG1 than mice sensitized with NG. By contrast, mice sensitized with NG produced higher levels of gliadin-specific IgG2a and INFγ. After a challenge, histamine levels were higher in mice sensitised with DG. DG can sensitize mice much more efficiently than NG. Moreover, this mouse model of allergy to DG revealed an IgE reactivity pattern against purified gliadins which was very similar to that of DG-allergic patients.

Modifying thioredoxin expression in cereals leads to improved pre-harvest sprouting resistance and changes in other grain properties

Thioredoxins (Trxs) are widely distributed, small proteins that function in redox regulation in a broad spectrum of cellular reactions. Experimental work with barley, wheat and a legume (Medicago truncatula) has established thioredoxin h (Trx h) as a central regulatory protein in seeds, reducing disulphide (S–S) groups of diverse seed proteins, including storage proteins, enzymes and enzyme inhibitors. Trxs appear to be particularly important in plants, as a large number of genes are present compared to mammalian organisms. In Arabidopsis thaliana there are approximately 20 different genes for classical Trxs, and large Trx gene families have also been found in cereals, such as rice, barley, wheat and sorghum. Extensive evidence indicates that adding Trx, reduced nicotinamide adenine dinucleotide phosphate (NADPH) and NADP-thioredoxin reductase (NTR) to cereal flour or seed preparations reduces disulphide (S–S) linkages of storage proteins. The early in vitro studies have been complemented with transgenic barley seed, overexpressing Trx h in protein bodies of the barley endosperm, which showed accelerated germination and early or enhanced expression of associated enzymes, i.e., α-amylase and pullulanase. Overexpression of Trx h levels in wheat was subsequently shown to (1) enhance protein solubility and digestibility, (2) reduce allergenicity of wheat gliadins, and (3) improve dough quality from poor-quality wheat flour. Most recently, we have demonstrated that down-regulation in wheat of Trx h9, a unique thioredoxin, leads to a reduction in the incidence of pre-harvest sprouting, demonstrated in several varieties over multiple generations with field-grown material. Yield and starch content were increased while baking quality in the high-gluten variety remained unchanged. These observations led to the intriguing question of how changes in the endosperm are communicated to the embryo. Studies of Trx h9, a membrane-associated Trx h that can move from cell to cell, provide suggestive evidence for a role of Trx h9 in intercellular communication of redox state.

Degradation of Wheat Epitope Peptides for Atopic Dermatitis and Exercise-Induced Anaphylaxis by Microbial Proteases

Epitope peptides of wheat gliadins and glutenins for atopic dermatitis, PQQPF and QQQPP, were degraded by the microbial proteases of Streptomyces griseus and Aspergillus oryzae. The proteases of Bacillus licheniformis and Bacillus amyloloiquefasiens degraded wheat gliadins and glutenins, but had little or no effect on the epitopes. A 16-mer peptide of CSQQQQPPFSQQQPPF, which incorporated this glutenin epitope twice, was also hydrolyzed by these proteases, and no peptide containing QQQPP was shown after the reaction with S. griseus protease. The proteases of S. griseus and A. oryzae also degraded epitope peptides of wheat gliadins and glutenins for exercise-induced anaphylaxis, QQIPQQQ, QQFPQQQ and QQPGQ.

Biochemical Properties of Bioplastics Made from Wheat Gliadins Cross-Linked with Cinnamaldehyde

The aim of this work has been to study the modification of gliadin films with cinnamaldehyde as a potential cross-linker agent. The molecular weight profile and cross-linking density showed that cinnamaldehyde increased reticulation in the resulting films. The participation of free amino groups of the protein in the newly created entanglements could be a possible mechanism of connection between the polypeptidic chains. The combination of a Schiff base and a Michael addition is a feasible approach to understanding this mechanism. The protein solubility in different media pointed to lower participation by both noncovalent and disulfide bonds in stabilizing the structure of the cross-linked films. The new covalent bonds formed by the cinnamaldehyde treatment hampered water absorption and weight loss, leading to more water-resistant matrices which had not disintegrated after 5 months. The properties of this novel bioplastic could be modified to suit the intended application by using cinnamaldehyde, a naturally occurring compound.

Conformational changes to deamidated wheat gliadins and β-casein upon adsorption to oil–water emulsion interfaces

The conformation of deamidated gliadins and β-casein in solution and adsorbed at the interface of oil-in-water emulsions was studied using synchrotron radiation circular dichroism (SRCD) and front-face-fluorescence spectroscopy. Deamidation led to partial unfolding of gliadins in solution. The α-helix content of the protein decreased from 35% (in the native form) to 16.3% while the percentage of β-sheet and unordered structure increased upon deamidation. The secondary structure of deamidated gliadins was largely unchanged upon adsorption to both tricaprin/water and hexadecane/water interfaces. In contrast, β-casein adopted a more ordered structure upon adsorption to these two oil/water interfaces, the α-helix content increased from 5.5% (in solution) to 20% and 22.5% respectively after adsorption to tricaprin/water and hexadecane/water interfaces. Both deamidated gliadins and β-casein have distinctive N-terminal hydrophilic and C-terminal hydrophobic domains. Unlike β-casein which contains no cysteine residue, gliadins have a large number of intramolecular disulphide bonds located in the C-terminal hydrophobic domain which constrains the conformational freedom of this protein upon adsorption to oil/water interfaces. The hydrophobicity of the oil phase also has an impact on the conformation of each protein upon adsorption to the oil/water interfaces – systematic trends were observed between oil phase polarity from: i) tryptophan fluorescence emission maxima, and ii) the α-helix content in the adsorbed state. Our results demonstrate that conformational re-arrangement of proteins upon adsorption to emulsion interfaces is dependent not only on the hydrophobicity of the oil phase, but more importantly on the conformational flexibility of the protein.

Wheat gluten fractions as wood adhesives—glutenins versus gliadins

AbstractPlant proteins, such as wheat gluten, constitute attractive raw materials for sustainable wood adhesives. In this study, alkaline water dispersions of the protein classes of wheat gluten, glutenin, and gliadin were used as adhesives to bond together wood substrates of beech. The aim of the study is to measure the tensile shear strength of the wood substrates to compare the adhesive performance of glutenin and gliadin and to investigate the influence of application method and penetration of the dispersions into the wood material. A sodium hydroxide solution (0.1M) was used as dispersing and denaturing agent. Dispersions with different protein concentrations and viscosities were used, employing wheat gluten dispersions as references. Two different application methods, a press temperature of 110°C and a press time of 15 min, were employed. The tensile shear strength and water resistance of the wood substrates were compared, using a slightly modified version of the European Standard EN 204. The bond lines of the substrates were examined by optical microscopy to study the penetration and bond‐line thickness. The results reveal that the adhesive properties of gliadin are inferior to that of both glutenin and wheat gluten, especially in terms of water resistance. However, the tensile shear strength and the water resistance of gliadin are significantly improved when over‐penetration of the protein into the wood material is avoided, rendering the adhesive performance of gliadin equal to that of glutenin and wheat gluten. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Wheat and maize allergy: which allergens are involved and relationship with symptoms severity

Cereals are the basis of human nutrition; they are the food source most intensively produced in the world, surpassing 2000 million tons annually. The production of wheat, corn and rice makes up over the 70% of the total cereal production and thus dominates world agriculture. All 3 cereal crops can determine adverse reactions with different mechanisms of immune-mediated hypersensitivity and through various routes of exposure (digestive, inhalation and contact). A paradigmatic example is that of wheat , which that can cause a range of reactions such as IgE-mediated food allergy, anaphylaxis and asthma, exercise-induced systemic reactions and cell-mediated reactions, such as celiac disease. The major allergens of cereals are represented by alpha-amylase inhibitors, some prolamins, such as gliadins in wheat and zeins in maize, and Lipid Transfer Proteins. These allergens are also differently implicated in the various clinical forms. So that their positivity in many cases actually can be considered diagnostic. The clinical diagnosis of allergy to cereals, because of their nutritional importance, should always be verified by double-blind placebo-controlled food challenged. Long term prognosis is quite favourable and the therapy is based essentially on their elimination, and on the treatment of symptoms in the case of ingestion.

Thermoplastic films from wheat proteins

We show that the wheat proteins gluten, gliadin and glutenin can be compression molded into thermoplastic films with good tensile strength and water stability. Wheat gluten is inexpensive, abundantly available, derived from renewable resource and therefore widely studied for potential thermoplastic applications. However, previous reports on developing thermoplastics from wheat proteins have used high amounts of glycerol (30–40%) and low molding temperature (90–120 °C) resulting in thermoplastics with poor tensile properties and water stability making them unsuitable for most thermoplastic applications. In this research, we have developed thermoplastic films from wheat gluten, gliadin and glutenin using low glycerol concentration (15%) but high molding temperatures (100–150 °C). Our research shows that wheat protein films with good tensile strength (up to 6.7 MPa) and films that were stable in water can be obtained by choosing appropriate compression molding conditions. Among the wheat proteins, wheat gluten has high strength and elongation whereas glutenin with and without starch had high strength and modulus but relatively low elongation. Gliadin imparts good extensibility but decreased the water stability of gluten films. Gliadin films had strength of 2.2 MPa and good elongation of 46% but the films were unstable in water. Although the tensile properties of wheat protein films are inferior compared to synthetic thermoplastic films, the type of wheat proteins and compression molding conditions can be chosen to obtain wheat protein films with properties suitable for various applications.

Immunological evaluation of the alcohol‐soluble protein fraction from gluten‐free grains in relation to celiac disease

Celiac disease (CD) is a gluten-sensitive enteropathy with an immune basis. We established the immune reactivity of the alcohol-soluble fraction from two minor cereals (tef and millet) and two pseudocereals (amaranth and quinoa) which are believed to be nontoxic based on taxonomy. Grains were examined in intestinal T-cell lines (iTCLs), cultures of duodenal explants from HLA-DQ2(+) CD patients and HLA-DQ8 transgenic mice for signs of activation. Our data indicated that tef, millet, amaranth, and quinoa did not show any immune cross-reactivity toward wheat gliadin, and therefore confirming their safety in the diet of CD patients.

Functional Properties of Bioplastics Made from Wheat Gliadins Modified with Cinnamaldehyde

Cinnamaldehyde is a naturally occurring α,β-unsaturated aldehyde. Its potential as a natural cross-linker to improve the physical performance of cast wheat gliadin films was evaluated. The cross-linking reaction was found to be dependent on the pH of the reaction medium, with pH 2 as the optimum. The water resistance (weight loss after immersion), mechanical properties (Young's modulus, tensile strength and elongation at break), thermal properties (T(g) and decomposition behavior), optical properties and morphology of films were evaluated. Cross-linked films showed high transparency, maintained their integrity after immersion, and displayed significant improvements in tensile strength and Young's modulus without impairment of their elongation properties. These effects, which were proportional to the amount of cinnamaldehyde added, highlight the possible formation of intermolecular covalent bonds between "monomeric" gliadins, leading to a polymerized network. Thus, this treatment could provide a new alternative to the toxic cross-linkers commonly employed and so extend the use of gliadin films.

Suppression of gliadins results in altered protein body morphology in wheat

Wheat gluten proteins, gliadins and glutenins, are of great importance in determining the unique biomechanical properties of wheat. Studies have therefore been carried out to determine their pathways and mechanisms of synthesis, folding, and deposition in protein bodies. In the present work, a set of transgenic wheat lines has been studied with strongly suppressed levels of γ-gliadins and/or all groups of gliadins, using light and fluorescence microscopy combined with immunodetection using specific antibodies for γ-gliadins and HMW glutenin subunits. These lines represent a unique material to study the formation and fusion of protein bodies in developing seeds of wheat. Higher amounts of HMW subunits were present in most of the transgenic lines but only the lines with suppression of all gliadins showed differences in the formation and fusion of the protein bodies. Large rounded protein bodies were found in the wild-type lines and the transgenic lines with reduced levels of γ-gliadins, while the lines with all gliadins down-regulated had protein bodies of irregular shape and irregular formation. The size and number of inclusions, which have been reported to contain triticins, were also higher in the protein bodies in the lines with all the gliadins down-regulated. Changes in the protein composition and PB morphology reported in the transgenic lines with all gliadins down-regulated did not result in marked changes in the total protein content or instability of the different fractions.

In vitro digestive stability of complexes between gliadin and synthetic blocking peptides

Celiac disease is caused by an inappropriate immune response to incompletely digested gluten proteins. We investigated whether synthetic peptides with high affinity to wheat gliadin could be selected with a phage display technique and whether complexes between such peptides and gliadin could sustain gastric and pancreatic digestion. Two synthetic peptides, P61 and P64, were selected because of their high affinity to immobilized gliadin. They were allowed to form complexes with gliadin, whereafter the complexes were subjected to in vitro digestion with gastric and pancreatic enzymes. The digestion products were analyzed with Western blot and RP HPLC. The results showed that both peptides formed stable complexes with intact gliadin and that complexes between gliadin and peptide P64 partly resisted gastrointestinal digestion. The two peptides reduced the binding of serum anti-gliadin IgA antibodies by 12%, and 11.5%, respectively, and the binding of anti-gliadin antibodies of the IgG isotype by 13% and 10%. Thus peptides produced by a phage display technique could interact stably with gliadin partly masking epitopes for antibody binding. A combination of peptides of this kind may be used to block gliadin-immune system interactions.

Glass transition of hydrated wheat gliadin powders

Modulated-temperature differential scanning calorimetric and dynamic mechanical analyses and dielectric spectroscopy were used to investigate the glass transition of hydrated wheat gliadin powders with moisture absorption ranged from 2.30 db% to 18.21 db%. Glass transition temperature (T g) of dry wheat gliadin was estimated according to the Gordon-Taylor equation. Structural heterogeneity at high degrees of hydration was revealed in dielectric temperature and frequency spectra. The activation energies (E a) of the two relaxations were calculated from Arrhenius equation.

Gliadin fragments promote migration of dendritic cells

In genetically predisposed individuals, ingestion of wheat gliadin provokes a T-cell-mediated enteropathy, celiac disease. Gliadin fragments were previously reported to induce phenotypic maturation and Th1 cytokine production by human dendritic cells (DCs) and to boost their capacity to stimulate allogeneic T cells. Here, we monitor the effects of gliadin on migratory capacities of DCs. Using transwell assays, we show that gliadin peptic digest stimulates migration of human DCs and their chemotactic responsiveness to the lymph node-homing chemokines CCL19 and CCL21. The gliadin-induced migration is accompanied by extensive alterations of the cytoskeletal organization, with dissolution of adhesion structures, podosomes, as well as up-regulation of the CC chemokine receptor (CCR) 7 on cell surface and induction of cyclooxygenase (COX)-2 enzyme that mediates prostaglandin E2 (PGE2) production. Blocking experiments confirmed that gliadin-induced migration is independent of the TLR4 signalling. Moreover, we showed that the α-gliadin-derived 31–43 peptide is an active migration-inducing component of the digest. The migration promoted by gliadin fragments or the 31–43 peptide required activation of p38 mitogen-activated protein kinase (MAPK). As revealed using p38 MAPK inhibitor SB203580, this was responsible for DC cytoskeletal transition, CCR7 up-regulation and PGE2 production in particular. Taken together, this study provides a new insight into pathogenic features of gliadin fragments by demonstrating their ability to promote DC migration, which is a prerequisite for efficient priming of naive T cells, contributing to celiac disease pathology.

Inhibition of angiotensin I-converting enzyme by wheat gliadin hydrolysates

A tryptic gliadin hydrolysate was fractionated into peptide fractions, which were assigned to either the central domain (CD) or terminal domains (TD) of gliadins. The domains were expected to contain amino acid (AA) sequences which, when released from the parent protein, inhibit the angiotensin I-converting enzyme (ACE), which plays a key role in regulating blood pressure. A proline (Pro) poor TD related fraction, containing the smallest peptides, showed the highest ACE inhibitory activity (IC 50 = 0.33 mg/ml). Additional peptidases were selected based on their in silico predicted ability to release ACE inhibitory peptides. Further hydrolysis of the tryptic hydrolysate fractions with thermolysin, Clarex, Alcalase and Esperase increased ACE inhibitory activities. Immobilised Ni 2+-ion affinity chromatography (IMAC) purification of a TD related peptide fraction obtained by sequential hydrolysis with trypsin and thermolysin yielded a fraction with an IC 50 value of 0.02 mg/ml. This IMAC fraction was enriched in histidine and hydrophobic AA (Pro, Val, Ile, Leu and Phe).

Effect of cleavage enzyme, search algorithm and decoy database on mass spectrometric identification of wheat gluten proteins

While tandem mass spectrometry (MS/MS) is routinely used to identify proteins from complex mixtures, certain types of proteins present unique challenges for MS/MS analyses. The major wheat gluten proteins, gliadins and glutenins, are particularly difficult to distinguish by MS/MS. Each of these groups contains many individual proteins with similar sequences that include repetitive motifs rich in proline and glutamine. These proteins have few cleavable tryptic sites, often resulting in only one or two tryptic peptides that may not provide sufficient information for identification. Additionally, there are less than 14,000 complete protein sequences from wheat in the current NCBInr release. In this paper, MS/MS methods were optimized for the identification of the wheat gluten proteins. Chymotrypsin and thermolysin as well as trypsin were used to digest the proteins and the collision energy was adjusted to improve fragmentation of chymotryptic and thermolytic peptides. Specialized databases were constructed that included protein sequences derived from contigs from several assemblies of wheat expressed sequence tags (ESTs), including contigs assembled from ESTs of the cultivar under study. Two different search algorithms were used to interrogate the database and the results were analyzed and displayed using a commercially available software package (Scaffold). We examined the effect of protein database content and size on the false discovery rate. We found that as database size increased above 30,000 sequences there was a decrease in the number of proteins identified. Also, the type of decoy database influenced the number of proteins identified. Using three enzymes, two search algorithms and a specialized database allowed us to greatly increase the number of detected peptides and distinguish proteins within each gluten protein group.

Foaming Properties of Wheat Gliadin

We studied gliadin solubility, surface tension and foam behavior, and the presence of different gliadin types in gliadin aqueous solutions and foams as a function of pH. Gliadin has excellent foaming properties only at neutral and alkaline pH. Its solubility is minimal near neutral pH, while almost complete at acidic and alkaline pH. Surface tensions of gliadin solutions are minimal around neutral pH, higher at alkaline pH, and highest at acidic pH, which corresponds well with their respective foaming properties. Foams at acidic and alkaline pH values are enriched in γ-gliadin, while foams at pH 8.0 have a similar distribution of α- and γ-gliadins. Thus, γ-gliadin predominantly contributes to the foaming properties of gliadin. The poor foaming properties of gliadin at pH 2.0 improve in the presence of 0.25 and 1.0% NaCl. It follows that the presence of positively charged amino acid residues hinders the formation of stable foam at acidic pH.

Deamidated gliadin antibodies

It was the discovery in 1997 that tissue transglutaminase (tTG) is the target autoantigen for anti-endomysial antibodies that triggered off significant progress both in the understanding of coeliac disease (CD) and in the serological assay available to assist in the diagnosis. The realisation that tTG is important in unmasking of B and T lymphocyte neo-epitopes by de-amidating glutamine residues to the more negatively charged glutamic acid was critical knowledge in this progress. Epitope mapping of de-amidated wheat gliadins and glutenins has uncovered some dominant linear B cell epitopes that have been employed in a number of novel assays now in routine use in diagnostic laboratories. These assays appear to have a similar diagnostic performance to the IgA anti-tTG/EMA assays with potential advantages in certain groups including those with IgA deficiency, young children and the elderly. How to best utilise de-amidated gliadin peptide (DGP) antibody assays remains an area of active debate and ongoing research. Given the complexity of wheat and other toxic CD proteins, research into B cell epitope mapping continues. This research has the potential discover more B cell epitopes and to further increase the sensitivity of DGP antibody assays without affecting specificity. This may make it the assay of choice for both screening and assisting in the diagnosis of CD in the future. It was the discovery in 1997 that tissue transglutaminase (tTG) is the target autoantigen for anti-endomysial antibodies that triggered off significant progress both in the understanding of coeliac disease (CD) and in the serological assay available to assist in the diagnosis. The realisation that tTG is important in unmasking of B and T lymphocyte neo-epitopes by de-amidating glutamine residues to the more negatively charged glutamic acid was critical knowledge in this progress. Epitope mapping of de-amidated wheat gliadins and glutenins has uncovered some dominant linear B cell epitopes that have been employed in a number of novel assays now in routine use in diagnostic laboratories. These assays appear to have a similar diagnostic performance to the IgA anti-tTG/EMA assays with potential advantages in certain groups including those with IgA deficiency, young children and the elderly. How to best utilise de-amidated gliadin peptide (DGP) antibody assays remains an area of active debate and ongoing research. Given the complexity of wheat and other toxic CD proteins, research into B cell epitope mapping continues. This research has the potential discover more B cell epitopes and to further increase the sensitivity of DGP antibody assays without affecting specificity. This may make it the assay of choice for both screening and assisting in the diagnosis of CD in the future.

Purified Wheat Gliadin Proteins as Immunoglobulin E Binding Factors in Wheat Mediated Allergies

Some wheat gliadin proteins are strong allergens that may cause various symptoms of food allergies and baker’s asthma. The most immunoreactive ω-5 gliadin fractions are the main allergens in wheat dependent exercise induced anaphylaxis (WDEIA). While the allergenicity of ω-5 is quite well understood, knowledge about α, β, γ and ω-1.2 gliadins is much more scanty. This study examines allergenic properties of other fractions as compared to ω-5. Gliadins were extracted from flour of winter wheat (Triticum aestivum L.) cultivar Ostka strzelecka. Purified samples representing proteins belonging to α, β, γ, ω-1.2 and ω-5 classes were isolated using preparative gel electrophoresis. Immuno-reactivity and allergenic properties of these proteins were analyzed by ELISA using sera from allergic patients with elevated sIgE (> 2KU/L), and by skin prick test (SPT). ELISA showed that ω-5 and ω-1.2 differed considerabely from α-, β- and γ-gliadins in respect of immunoreactivity. Responses of both ω-gliadins were almost twice as high as for other fractions. Significant differences were also observed among individual ω-gliadin fractions as evidenced by ANOVA. SPT showed that patient with symptoms of baker’s asthma and WDEIA had a positive results to all gliadins tested. Another patient with baker’s asthma (but not WDEIA) reacted positively only to ω-5 gliadins. In two patients with skin allergy SPT were negative with all analyzed proteins. Results show ω-1.2 gliadins to be almost as immunorective as ω-5. The α-, β- and γ-gliadins also recognize specific IgE antibodies, but their binding capacity is only about half that of ω-fractions. This kind of immunoreactivity could still be important since a cumulative effect of individual fractions may intensify disease symptoms in allergic patients.

Ages of celiac disease: From changing environment to improved diagnostics

From the time of Gee's landmark writings, the recent history of celiac disease (CD) can be divided into many ages, each driven by a diagnostic advance and a deeper knowledge of disease pathogenesis. At the same time, these advances were paralleled by the identification of new clinical patterns associated with CD and by a continuous redefinition of the prevalence of the disease in population. In the beginning, CD was considered a chronic indigestion, even if the causative food was not known; later, the disease was proven to depend on an intolerance to wheat gliadin, leading to typical mucosal changes in the gut and to a malabsorption syndrome. This knowledge led to curing the disease with a gluten-free diet. After the identification of antibodies to gluten (AGA) in the serum of patients and the identification of gluten-specific lymphocytes in the mucosa, CD was described as an immune disorder, resembling a chronic "gluten infection". The use of serological testing for AGA allowed identification of the higher prevalence of this disorder, revealing atypical patterns of presentation. More recently, the characterization of autoantibodies to endomysium and to transglutaminase shifted the attention to a complex autoimmune pathogenesis and to the increased risk of developing autoimmune disorders in untreated CD. New diagnostic assays, based on molecular technologies, will introduce new changes, with the promise of better defining the spectrum of gluten reactivity and the real burden of gluten related-disorders in the population. Herein, we describe the different periods of CD experience, and further developments for the next celiac age will be proposed.