Prospective study of sensitization and food allergy to flaxseed in 1317 subjects.

BACKGROUND: Foods containing flaxseed proteins rich inpolyunsaturatedfatty acids are new on the market. OBJECTIVES: In a population of patients attending the allergology department, we evaluated the frequency of sensitization to flaxseed, characterized allergens and looked for modifications related to industrial processing. METHODS: Natural, heated and extruded flaxseeds were tested using prick-in-prick tests (PIP using the fresh seed), SDS PAGE, immunoblots, immunoblot inhibition and Fourier Transform Infrared (FTIR) spectroscopy. RESULTS: PIP tests to natural flaxseed were positive in 5.8% of the 1317 patients. 73 of 77 PIP-positive patients were atopic. There was cross-reactivity with five seeds. peanut, soybean, rapeseed, lupine and wheat, and with rape pollen. Immunoblot inhibition by bromelain confirmed the presence of specific IgE to cross-reactive carbohydrate determinants (CCD). 0.15% of this population presented with food allergy to flaxseed and positive PIP to heated and extruded flaxseed. Two sera showed that clinically relevant allergens in industrial products had MW between 25 and 38 kDa. Sensitization to processed flaxseed characterized only the allergic subjects. FTIR spectroscopy showed major modifications in 3 and alpha structures following industrial processing. CONCLUSION: Positive prick tests to natural flaxseed were mainly due to cross-reactions. Flaxseed allergy is rare and could be detected by PIP to heated extruded flaxseed. Increasing consumption callsfor monitoring of clinical risk.

In vitro allergenicity of peanut after hydrolysis in the presence of polysaccharides.

BACKGROUND: Peanut is a major allergenic product. Manufacturing processes used in food industries to improve the physicochemical properties of food-based peanut (stabilization, texturization), could cause a modification of the digestibility of peanut proteins and, consequently, their allergenicity. OBJECTIVE: This study aimed at examining the influence of polysaccharides, i.e., gum arabic, low methylated pectin (LMP) and xylan, on the in vitro hydrolysis of peanut protein isolate (PPI) and the in vitro allergenicity of the digestion products. METHODS: PPI was hydrolysed during a two-step in vitro hydrolysis by pepsin, followed by a trypsin/chymotrypsin (T/C) mixture performed in dialysis bags with molecular weight cut-offs (MWCO) of 1000 or 8000 Da. SDS-PAGE electrophoresis and immunoblotting were assessed on the peptic and T/C digestion products in (retentates) and out of the dialysis bags (dialysates). RESULTS: Hydrolysis by all of the digestive enzymes showed retention of some proteins in the dialysis bags in the presence of gum arabic and xylan. The retentates were recognized by IgG and IgE, particularly peptides <20 kDa. The IgE binding with peptides of retentate containing xylan from the dialysis bag with an MWCO of 1000 Da was reduced. The immunoreactivity of hydrolysis products in dialysates was considerably reduced by polysaccharides, regardless of the dialysis bag. CONCLUSION: Reduction of PPI hydrolysis was probably due to non-specific interactions between polysaccharides and peptides. In retentates, IgE-binding epitopes were reduced by digestion and the presence of xylan. In dialysates, they were reduced by all of the polysaccharides. This work highlights the possibility of modulating this food allergy through optimized formulation.

[Food allergy: effect of proteins-lipids and proteins-polysaccharides interactions]. / Allergénicité alimentaire: importance des interactions protéines-lipides et protéines-polysaccharides.

The most widely used ingredients in food formulation are proteins, lipids and polysaccharides. Proteins-lipids and proteins-polysaccharides interactions play a key role in the structure, stability, sensorial and nutritional properties of formulated foods. The objective of the present study is to highlight the importance of proteins-lipids and proteins-polysaccharides interactions, on the immuno-reactivity of allergenic proteins. Two models have been studied, on the one hand refined and not refined oils (soya and sunflower) and soya lecithin, on the other hand mixtures based on peanut proteins and polysaccharides (arabic gum, pectin, xylan). STUDY OF OILS: We have extracted proteins, using a PBS buffer, from refined and not refined oils from soya, sunflower and from soya lecithin, determined protein concentrations and identified allergenic proteins using SDS-PAGE electrophoresis and immuno-blotting. Phospholipids are determined by atomic absorption spectrometry. The protein determination and SDS-PAGE show the presence of a higher amount of proteins in not refined oils and lecithin as compared to refined oils. An important amount of proteins associated to phospholipids are eliminated by degumming on the form of lecithin. On the other hand, residual proteins from refined oils are accompanied by phospholipids. Immuno-blots reveal the presence of a 56 kDa allergen in oils issued from soya seeds and soya lecithin, and the presence of a 67 kDa allergen in oils issued from sunflower seeds. We conclude that the presence or elimination of proteins, especially allergens from oils is linked to amphiphilic association to phospholipids. STUDY OF PEANUT PROTEINS-POLYSACCHARIDES MIXTURES: We have digested in vitro proteins in a dialysis bag using a multi-enzymatic method and characterized proteins and peptides using SDS-PAGE electrophoresis and immuno-blotting. Our results confirm that peanut proteins alone are digested by proteases and that a number of large peptides still have epitopes recognized by anti-peanut proteins antibodies. Our results also show that the presence of polysaccharides changes the peptidic profile after digestion and that, depending on the polysaccharide type, smaller or larger peptides can be obtained in the dialysis bag. Smaller peptides are obtained using pectin whereas larger peptides are obtained using arabic gum and xylan. In the latter case, an increasing amount of peptides reacts to antibodies. Our first observations clearly show the need to better understand modifications of proteins allergenicity induced by the presence of other ingredients such as polysaccharides and lipids, in relation to technological treatments.

Effects of different levels of gum arabic, low methylated pectin and xylan on in vitro digestibility of beta-lactoglobulin.

Plant hydrocolloids used in the food industry to improve texture and stability of food, such as dairy products, can reduce protein digestibility and, consequently, modify the bioavailability of amino acids. We studied the in vitro hydrolysis at 37 degrees C of beta-lactoglobulin (beta-lg) in mixed dispersions containing either gum arabic or low-methylated pectin or xylan at levels of 0, 1, 10, 20, 30, and 50% weight. Proteolysis used either pepsin alone by progressive reduction of pH during proteolysis or pepsin followed by trypsin and chymotrypsin in two different dialysis bags with a molecular weight (MW) cutoff of 1000 or 8000 Da. Results showed that beta-lg was almost resistant to pepsin digestion and that the three plant hydrocolloids inhibited significantly beta-lg digestibility as determined using dialysis bag with a 1000-Da MW cutoff. Among the three polysaccharides used, xylan showed a digestibility decrease greater than that obtained with gum arabic and low-methylated pectin. On the other hand, no significant effect of polysaccharides on the in vitro beta-lg digestibility was detected using the dialysis bag with an 8000 Da MW cutoff. This mainly suggests that peptides with MW in the range 1000 to 8000 Da may interact with polysaccharides more than peptides and proteins with a greater molecular weight to decrease the protein digestibility, and that the nature of the polysaccharides plays a role in the interaction.

[Biological exploration of food allergy]. / Exploration biologique de l'allergie alimentaire.

Most of the time, food allergies are the consequence of an immediate IgE mediated hypersensitivity. In France, the frequency of food allergies is of 3.24%, it raises 8% in children. Their number as been multiplied by 2 in 5 years. The biological approach of food allergy is based essentially on the detection and the measurement of specific IgE. To date, a high number of tests are marketed and it is important to determine which of them are of high quality, using analytical or clinical-biological evaluations. Some other ways are available to study thoroughly and to characterize the allergens responsible of clinical reactions: the detection of hidden allergens, the study of crossed reaction between food or between food and pollen, the modification of the allergens by food processing.

[Allergenicity of oils]. / Allergénicité des huiles.

Cases of allergy to the oils of groundnut, sunflower, soya and sesame have been described in the literature. In parallel, other authors have affirmed that these oils are not allergenic. The objective of this article is to make the point on this question, to cite the procedures to which the seeds are submitted to extract the oil, to remember that the oils are not composed only of triglycerides and to describe the results of our work. Allergy of oils is a subject that is constantly submitted to controversy and the bibliography does not cease to give contradictory examples. This may be explained by the variations in extraction procedures used by the manufactures, as well as by the conditions of extraction of the proteins in the laboratory.

Influence of refining steps on trace allergenic protein content in sunflower oil.

BACKGROUND: Although allergy to sunflower seed and oil is a relatively rare occurrence, several cases of sunflower seed allergy have been observed, and we have already described one case of anaphylaxis after eating sunflower oil and margarine. OBJECTIVE: The aim of our study was to determine and characterize the allergens from sunflower oil at the different steps of the refining process: crude pressed oil (step A), acidification and neutralization (step B), pregumming by centrifugation (step C), washing (step D), bleaching (step E), gumming by filtration (step F), and deodorization (step G). METHODS: A sample of oil from each step of the process (steps A to G) was heat extracted with PBS. The protein concentration of each extract was evaluated by using the micro-Bradford assay. Samples were run on SDS-PAGE. The immunoblot was performed with the serum of a patient sensitized to sunflower seed and oil. RESULTS: The extracts obtained after each step reveal a decrease in total protein concentration from 13.6 microg/mL to 0. 22 microg/mL. The result of SDS-PAGE shows 5 bands, from 67 kd to 145 kd, with the most abundant being the 67-kd protein. The amount of this protein decreases after each step of the process. It is, however, still present in trace amounts in the refined oil. The 67-kd protein, which is mainly present in the crude oil and slightly in the refined oil, has been shown to be allergenic. CONCLUSION: Because of the presence of allergenic proteins, refined sunflower oil may pose a threat to people highly sensitized to sunflower seeds.

Cross-allergenicity of peanut and lupine: the risk of lupine allergy in patients allergic to peanuts.

BACKGROUND: Peanut allergy is common, but cross-allergy between legumes is rare. Proteins from Lupinus albus are increasingly eaten in the form of seeds or additives to wheat flour. The risk of cross-allergenicity is still insufficiently known. OBJECTIVE: We sought to study the risk of cross-allergy to lupine in patients allergic to peanut and to study lupine allergenicity. METHODS: Twenty-four patients allergic to peanuts were studied by means of skin prick tests with native lupine flour from Lupinus albus. Double-blind oral challenge tests were performed with lupine flour and peanut in 8 of these patients. Specific IgEs were assayed for peanut, lupine flour, and pollen in 6 sera. RAST inhibition tests for lupine pollen by peanut were performed on 4 of these sera. Peanut and lupine flour immunoblots were carried out for 6 sera, and crossed immunoblot inhibitions for peanut by lupine flour and lupine flour by peanut were carried out for 2 sera. RESULTS: The skin prick test responses with lupine flour were positive in 11 (44%) subjects. The challenge test responses were positive in 7 of 8 subjects at the same doses as with peanut. The major lupine flour allergen (molecular mass, 43 kd) is present in peanuts. The RAST inhibition and immunoblot tests indicated cross-reactivity of peanut with the lupine flour and pollen. CONCLUSIONS: The risk of crossed peanut-lupine allergy is high, contrary to the risk with other legumes. The inclusion of 10% lupine flour in wheat flour without mandatory labeling makes lupine a hidden allergen, presenting a major risk of cross-reaction in subjects already allergic to peanut products. A high sensitizing potential can also be postulated for this legume.

Anaphylaxis to mustard as a masked allergen in "chicken dips".

BACKGROUND: Masked allergens in processed foods are sometimes responsible for "idiopathic" anaphylaxis because of the lack of full ingredient labeling. Patients, even if they are aware of a specific hypersensitivity, may be unaware of trace amounts of the offending allergen(s) that are present in these products. METHODS: We report a case of severe anaphylaxis to mustard masked in "chicken dips." A 38-year-old woman had an anaphylactic reaction beginning 20 minutes after eating "chicken dips" in a fast food restaurant. She was treated successfully in an emergency room and then referred to us for evaluation. We performed skin prick tests, IgE antibody, and inhibition assays to identify allergens in foods eaten just prior to the anaphylactic reaction. RESULTS: She had a history of allergy to mustard. Skin prick tests revealed sensitivity to mustard, coriander, and curry powder, which contains mustard. She had 56.3 KU/L mustard-specific serum IgE antibody in a Phadebas CAP RAST assay (Pharmacia, Sweden). The mustard RAST was inhibited with an extract of "chicken dips," confirming the presence of mustard in "chicken dips." CONCLUSIONS: This case points out the risk of masked allergens in modern food products and the value of the RAST inhibition assay for the identification of such hidden allergens.