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IgA and IgG immunoreactivity of multiple myeloma patients sera with gliadin from diiferent wheat cultivar.
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Immunity to food antigens (gliadin, cow's milk proteins) is in the centre of the attention of modern medicine focused on the prevention of diseases, prevention which is based on the use of appropriate restriction diet. Detection of the enhanced levels of the immune reactions to antigen(s) present in food is from this point of view of great importan...
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... If this situation is related to the local wheat varieties and characteristic food in Xinjiang? Wheat varieties in Xinjiang are rich in genetic diversity (Wei et al., 2001), and previous studies had shown that the contents of T-cell stimulatory epitopes vary among diverse wheat cultivars (Konic-Ristic et al., 2009;Van den Broeck et al., 2010a). This finding opens possibilities for the selection of wheat varieties with low levels of CD epitopes. ...
Xinjiang is a high‐risk area for celiac disease (CD) regardless of genetic or environmental factors. However, no case has been reported yet in people living in Xinjiang. This study aims to explore the potential connection between diet and occurrence of CD in the Xinjiang population. To this end, the levels of T‐cell stimulatory epitopes in 164 accessions of Xinjiang wheat were tested by using Western blot with monoclonal antibodies against α‐gliadin epitopes Glia‐α9 and Glia‐α20. Three wheat varieties with remarkably low amounts of T‐cell stimulatory epitopes were obtained. Western blot and R5 competitive ELISA were performed for the assessment of potential toxicity related to CD of naan. Results showed a reduction of gluten toxicity after wheat flour was processed into naan, suggesting it may have the potential to help to reduce the risk of CD for the genetically predisposed individuals.
... Celiac disease is a gluten sensitive disease [4]. Gluten is derived from wheat, barley, and rye [5]. ...
... Barley flour was used as a positive control, and 11 of 31 active-CD patients demonstrated significant antibody binding to the barley extract. Although there are some identical gluten peptides found in wheat and barley, cultivated barley has been shown to be less immunogenic than wild-type barley (52), and celiac patients have variable responses to different varieties of barley and wheat (54), so these results were not unexpected. In addition, the ELISAs were performed on native proteins and not on proteins deamidated by tTG, which might enhance their immunoreactivity. ...
Enzymatic digestion, or hydrolysis, has beenproposed for treating gluten-containing foods andbeverages to make them safe for persons with celiacdisease (CD). There are no validated testing methodsthat allow the quantitation of all the hydrolyzed orfermented gluten peptides in foods and beverages thatmight be harmful to CD patients, making itdifficult to assess the safety of hydrolyzed products.This study examines an ELISA-based method todetermine whether serum antibody binding ofresidual peptides in a fermented barley-basedproduct is greater among active-CD patients thana normal control group, using commercial beersas a test case. Sera from 31 active-CD patients and29 nonceliac control subjects were used to assessthe binding of proteins from barley, rice, traditionalbeer, gluten-free beer, and enzymatically treated(gluten-removed) traditional beer. In the ELISA,none of the subjects' sera bound to proteins in thegluten-free beer. Eleven active-CD patient serumsamples demonstrated immunoglobulin A (IgA) orimmunoglobulin G (IgG) binding to a barley extract,compared to only one nonceliac control subject. Ofthe seven active-CD patients who had an IgA bindingresponse to barley, four also responded to traditionalbeer, and two of these responded to the glutenremovedbeer. None of the nonceliac control subjects'sera bound to all three beer samples. Binding ofprotein fragments in hydrolyzed or fermented foodsand beverages by serum from active-CD patients,but not nonceliac control subjects, may indicate thepresence of residual peptides that are celiac-specific.
... However, a recent paper demonstrated that common wheat, rye, and barley cultivars varied in this ratio, and the gluten content will be overestimated using the general conversion factor (WIESER & KOEHLER, 2009). Many studies have focused on the diversity of the celiac-toxic protein profi le of cereals, and have shown that there are differences in the immunogenic prolamin content not only among wheat, barley, and oat species but within cultivars, too (KONIC-RISTIC et al, 2009;COMINO et al., 2011COMINO et al., , 2012PRANDI et al., 2012). This diversity relates to different expression patterns of storage proteins, which is primarily determined genetically, but it is also infl uenced by environmental factors, such as soil composition, weather, infections, heat or cold shock (DUPONT & ALTENBACH, 2003;WIESER et al., 2004). ...
In special dietary products for people intolerant to gluten, gluten content is not supposed to exceed the regulatory thresholds. Enzyme-linked immunosorbent assays (ELISAs) are routinely used to quantitate gluten in these products. They measure gliadin/gluten with high specificity and sensitivity, but they have some limitations. Quantitative and qualitative variability of the target proteins among wheat cultivars is a factor that may cause inaccurate results. One of the aims of this work was to characterize the protein composition of five wheat cultivars grown in multiple harvest years and their blends by reversed-phase high-performance liquid chromatography (RP-HPLC). The gliadin/gluten content of these wheat flours was also analysed with two commercial ELISA kits. The effect of differences in protein profiles between the flours from an individual cultivar and the blend of five cultivars, harvest years, as well as processing procedures (dough forming and baking) on the results of two ELISA kits was investigated and their relative magnitude was determined. Among the factors investigated, the differences between flours had the greatest impact on gliadin recoveries.
... Analysis of gliadin and glutenin immunogenicity. IgA and IgG immunoreactivity to the same protein content of crude gliadins (Sigma-Aldrich, USA), as well as gliadins and glutenins extracted from non-treated, steeped and sprouted wheat grains was investigated by using home-made ELISA as described by Konić-Ristić et al. 15 The level of specific immunoglobulins of the IgA-and IgG-type was determined by measuring the absorbance after adding the secondary antibody (anti-IgA/anti-IgG, 1 : 2500) labeled with peroxidase and a suitable substrate (TMB, 3,3′,5,5′-tetramethylbenzidine). The absorbances were measured at 450 nm. ...
The aim was to determine the effect of steeping and sprouting on wheat grain proteins and the functional
consequences in this regard. The solubility of proteins and the polypeptide composition of albumins, globulins,
gliadins and glutenins were determined, as well as the content of non-protein nitrogen and free
sulfhydryl groups (–SH), and the activity of peroxidase (POD) and lipoxygenase (LOX). In addition, the
pasting viscosity of flour and protein bioactivity such as antioxidant capacity and immunoreactivity were
evaluated. The increase of non-protein nitrogen and free –SH groups by about 62.09 and 96.7%, respectively,
as well as the decrease of albumin + globulin polypeptides with a molecular weight over 85.94 kDa
and between 85.94–48.00 kDa by about 34 and 8.7%, respectively, were the most notable changes
observed in the flour from whole sprouted wheat that clearly point to the intensive protein hydrolysis. The
reduction of disulfide bonds and increased concentrations of free –SH groups significantly modify the
visco-elastic properties of gliadins and glutenins causing pasting viscosity reduction. However, sprouted
wheat flour could be considered as a potential food ingredient because of its improved antioxidant
capacity that is a result of protein hydrolysis inter alia. As protein modification induced by steeping may
have beneficial effects on the antigenicity of the glutenin fraction, this kind of wheat pretreatment can
represent a putative strategy in the dietary modulation of diseases related to glutenin immunoreactivity,
e.g. dermatitis herpetiformis.
... The ELISA assay is based on a monoclonal antibody R5, which recognizes epitopes of amino acid sequences QQPFP, QQQFP, LQPFP, and QLPFP in wheat gliadins and corresponding proteins from barley (hordeins) and rye (secalins) (Kahlenberg et al., 2006;Konic-Ristic et al., 2009;Van Eckert et al., 2010). Although R5 antibody is declared to have no cross-reactivity with oat proteins, avenins may contain QQQPF sequences and they can also react properly with R5 antibody (Comino et al., 2011;Ellis et al., 1998;Osman et al., 2001). ...
Background. The aim of this study was to compare the biochemical and immunochemical properties of avenins in some special oat raw materials and additionally the possibility of using them as a raw material for the gluten-free bakery products.
Materials and methods. The compared oat raw materials were – oat flakes, commercial oat flours (includ- ing gluten-free oat flour) and residual oat flour, which is by-product of β-glucan preparation. Biochemical characteristic included amino acid compositions and SDS-PAGE profiles of extracted avenins. The immuno- chemical reactivity with polyclonal anti-gluten and monoclonal anti-gliadin antibodies was evaluated quali- tatively and quantitatively by immunoblotting and ELISA methods. Additionally, experimental bakery prod- ucts made of examined raw materials were assessed according to their suitability for the celiac patients’ diet. Results. The highest protein content was measured in the β-glucan preparation “Betaven“ and gluten-free oat flour. Proteins of all materials are rich in glutamic and aspartic acid, leucine and arginine. Proportions of amino acids in avenins extracted from most of oat raw materials are similar, excluding gluten-free oat flour, which has a very low avenin content and proportions of individual amino acids are different. The SDS-PAGE protein pattern consisted of proteins with molecular weight of about 25–35 kDa. Polyclonal anti-gluten anti- body recognized all protein fractions of molecular weight higher than 20 kDa. Quantitative ELISA analysis shows that the majority of samples has a gliadin-like protein content within the range of 80–260 mg/kg, excluding gluten-free flours and corresponding bakery products. Altogether, β-glucan preparation has ex- tremely high level of gliadin-like proteins.
Conclusion. In the examined oat raw materials and foods the contents of immunoreactive amino acid se- quences exceeded the limit of 20 mg/kg (considered as gluten-free) except for gluten-free flours (oat and the prepared mixture) and the bakery products based on gluten-free flours. Unfortunately, the rest of oat raw materials and products cannot be considered gluten-free.
... In our previous articles we proved that many of food antigens (like gliadin from wheat, cow's milk proteins, phytohemagglutinin from red beans), induced some kind of immune-mediated molecules -synthesis of various class of immunoglobulins, IgG, IgA, IgM [19][20][21]. Similarly to this it was easy to propose that the edible mushrooms as the rich source of tyrosinase and of melanin, after the consummation, could induce immunity to mentioned molecular structures. ...
The aim of this study was to determine the presence and the intensity of humoral immunity to melanoma-associated antigens: tyrosinase and melanin, in patients with melanoma, in persons with vitiligo and in control healthy people.
The study involved 63 patients with melanoma and 19 persons with vitiligo. Control group consisted up to 41 healthy volunteers. Mushroom tyrosinase and synthetic melanin were used as the antigens.
ELISA test showed significantly (p < 0.0000004 and p < 0.04) lower levels of IgM anti-tyrosinase autoantibodies, in melanoma and vitiligo patients respectively, compared to controls.Although there was no significant difference between the levels of IgA anti-melanin autoantibodies in melanoma or vitiligo patients in comparison with controls, the enhanced concentrations of anti-melanin IgA autoantibodies were preferentially found in melanoma patients with metastatic disease. Significantly high percentage in the Fc alphaRI (CD89) positive cells was determined in melanoma patients (p < 0.002 and p < 0.008) in comparison to that found in healthy people or in patients with vitiligo, in the already mentioned order, pointing that IgA dependent cellular cytotoxicity is not important for the immune action against melanoma, even more that it is included in some immune suppression.Levels of IgG autoantibodies to mentioned antigens in melanoma patients although low were not significantly lower from controls. These findings analyzed together with the statistically significant low percentage of FcgammaRIII, (CD16) positive immunocompetent cells (p < 0.0007 and p < 0.003), which was found in patients with melanoma compared with healthy or vitiligo people respectively, and statistically significant low percentage of (CD16 + CD56+) natural killer (NK) cells (p < 0.005) found in melanoma patients in comparison to healthy controls pointed to the low probability for anti-melanoma IgG mediated, antibody mediated cellular cytotoxicity, (ADCC) and NK cytotoxicity. Moreover the ratio of the percentages of granulocytes and percentage of lymphocytes was statistically higher in patients with melanoma in relation to healthy people as well as to people with vitiligo (p < 0.0007 and p < 0.05 respectively).
Autoantibodies to tyrosinase and to melanin which are found even in healthy people, point that consummation of edible mushrooms that carry the antigen tyrosinase and melanin, could influence the humoral anti-melanoma immune response.Levels of different immunoglobulin classes of anti-melanin and anti-tyrosinase antibodies varied depending on the presence and the stage of studied diseases. Besides, the statistically enhanced ratio of the percentages of granulocytes and percentage of lymphocytes, together with statistically decreased percentage of NK cells is found in analyzed melanoma patients.
... Several monoclonal or polyclonal anti-gliadin antibodies were developed for immunological tests for determination of gliadin content, e.g. polyclonal antibodies developed against wheat gliadin, or a monoclonal antibody made against w-gliadins (Ristic et al. 2009 ). At the present time the greatest attention has monoclonal antibody R5, developed against a secalin extract, which recognizes the epitopes with the amino acid sequences QQPFP, QQQFP, LQPFP and QLPFP (Kahlenberg et al. 2006; van Eckert et al. 2010). ...
... At the present time the greatest attention has monoclonal antibody R5, developed against a secalin extract, which recognizes the epitopes with the amino acid sequences QQPFP, QQQFP, LQPFP and QLPFP (Kahlenberg et al. 2006; van Eckert et al. 2010). These epitopes occur repeatedly to a similar level in a-, g-and w-gliadins, hordeins and secalins of wheat, barley and rye (Konic-Ristic et al. 2009). In this study, we analysed the prolamin proteins of cereal and alcohol soluble proteins of pseudocereal and legume grains by comparison of protein fractions, amino acids composition , SDS-PAGE electrophoresis and immunoreactivity tested by Western blot and ELISA methods. ...
Prolamins are alcohol-soluble fraction of cereal proteins involved in immunological response
of patients with celiac disease. The aim of this study was to analyse the similar protein
complex of selected varieties of cereal, pseudocereal and legume grains by comparison of protein
fractions, amino acids composition and SDS-PAGE electrophoresis. The immunoreactivity
was tested by Western blot and ELISA methods. ELISA analysis recognizes celiac active
epitopes in wheat gliadin (which is reference protein in determination of celiac activity),
and also corresponding epitopes in other grain proteins responsible for immunological response
of patients with celiac disease. Estimated quantity of celiac active sequences (calculated
as gliadin content) below 20 ppm was found in species of amaranth, buckwheat and millet,
as well as rice, maize, chickpea and chickling vetch. Immunological reaction with
polyclonal antibody was negative for all crops, except oat, maize, millet and foxtail millet.
Food-derived opioid peptides include digestive products derived from cereal and dairy diets. If these opioid peptides breach the intestinal barrier, typically linked to permeability and constrained biosynthesis of dipeptidyl peptidase-4 (DPP4), they can attach to opioid receptors. The widespread presence of opioid receptors spanning gut, brain, and internal organs is fundamental to the diverse and systemic effects of food-derived opioids, with effects being evidential across many health conditions. However, manifestation delays following low-intensity long-term exposure create major challenges for clinical trials. Accordingly, it has been easiest to demonstrate causal relationships in digestion-based research where some impacts occur rapidly. Within this environment, the role of the microbiome is evidential but challenging to further elucidate, with microbiome effects ranging across gut-condition indicators and modulators, and potentially as systemic causal factors. Elucidation requires a systemic framework that acknowledges that public-health effects of food-derived opioids are complex with varying genetic susceptibility and confounding factors, together with system-wide interactions and feedbacks. The specific role of the microbiome within this puzzle remains a medical frontier. The easiest albeit challenging nutritional strategy to modify risk is reduced intake of foods containing embedded opioids. In future, constituent modification within specific foods to reduce embedded opioids may become feasible.
Differences in the level of coeliac-active gluten epitopes in wheat might have some significance for individuals reporting noncoeliac gluten sensitivity. The aim of this study was to compare the reactivity of epitopes towards ELISA R5 and G12 monoclonal antibodies in ancient (emmer; Khorasan wheat; spelt) and modern wheat (common bread wheat; durum), and to check whether the bread-making process leads to the degradation of epitopes. Data from ELISA R5 and G12 did not match gluten dry weight in wheat. Bread dough fermentation and extensive baking did not change the reactivity of coeliac-active epitopes towards monoclonal antibodies. Compared to hexaploid bread-type wheat (spelt; common bread wheat), ancient and modern pasta-type tetraploid wheat (emmer; Khorasan; durum) had less epitopes reactive towards ELISA R5 and G12 and might be preferable for wheat-sensitive individuals looking for food with reduced coeliac-active epitopes.
