muramidase and Food-Hypersensitivity

The Drug Allergy Committee of the Spanish Society of Allergology and Clinical Immunology reviewed the allergenic potential of several substances of food origin that are found in the composition of some drugs. Despite recent legislation on labeling, many labels do not clearly state whether the drug contains raw material (active ingredients, excipient, or other manufacturing intermediate) with an origin in any of the substances in the list of the 14 groups of food allergens that are subject to mandatory declaration. The objective of legislation is that the drug package, the Summary of Product Characteristics, and the patient information leaflet clearly state the food content in order to improve the safety of allergic patients. Therefore, any food or allergen derivative that must be declared should be clearly stated on the drug label. Of all the evaluated products, egg and milk derivatives are the most frequently discussed in literature reviews. The natural or synthetic origin of potentially allergenic substances such as lysozyme, casein, lactose, albumin, phosphatide, and aromatic essences should be clearly stated. Providing this information has 2 clear advantages. First, allergic reactions to drugs in patients with food allergy could be avoided (if the substances have a natural origin). Second, prescription would improve by not restricting drugs containing synthetic substances (which do not usually induce allergic reactions).

Topics: Drug Hypersensitivity; Food Additives; Food Hypersensitivity; Glucosamine; Humans; Lactose; Muramidase; Ovalbumin; Propofol; Spain

Topics: Allergens; Amino Acid Sequence; Animals; Cattle; Cross Reactions; Dietary Proteins; Food Hypersensitivity; Humans; Lactalbumin; Lactoglobulins; Molecular Sequence Data; Muramidase; Proteins; Serum Albumin

We occasionally see egg-allergic children who develop contact urticaria to hen's egg despite the absence of the overt symptoms on ingestion. The mechanisms remain to be elucidated.. Twenty-one subjects with positive reactions to 20-min patch tests for egg-white antigens were divided into subgroups with positive (n = 10) and negative (n = 11) results to oral challenge tests by the same antigens. We measured IgE antibody for egg white and its components, and IgE-binding activities to digestive enzyme-treated ovomucoid by RAST inhibition.. There were no significant differences in IgE antibody titers to egg white (positive vs negative: 30.3% vs 15.3%, P=0.130), ovomucoid (21.5% vs 10.2%, P= 0.078), ovotransferrin (9.9% vs 3.7%, P = 0.105), and lysozyme (3.4% vs 2.9%, P=0.944), except ovalbumin (16.8% vs 5.6%, P=0.024), between the positive and negative subjects in the provocation tests. In contrast, the concentration (1.93 microg/ml) of pepsin-treated ovomucoid needed for 50% RAST inhibition in the challenge-positive subjects was significantly (P=0.0003) lower than that (114.9 microg/ml) of negative subjects. Similar but less significant differences were obtained when ovomucoid fragments treated with chymotrypsin (0.91 microg/ml vs 6.86 microg/ml, P=0.014) and trypsin (0.75 microg/ml vs 4.67 microg/ml, P= 0.041) were used as inhibitors.. We suggest that IgE antibodies from subjects showing contact urticaria despite the absence of reactions to the ingestion of egg white recognize the epitope(s) unstable to digestive enzymes.

Topics: Administration, Oral; Allergens; Animals; Child; Child, Preschool; Chymotrypsin; Conalbumin; Double-Blind Method; Egg White; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Male; Muramidase; Ovalbumin; Ovomucin; Pepsin A; Placebos; Radioallergosorbent Test; Trypsin; Urticaria

No egg white products have been clearly proven to be hypoallergenic. The role of egg white proteins in allergic reactions to eggs is still debatable.. This study was designed to determine the importance of ovomucoid, an egg white protein, in the development of allergies to egg white.. We performed a double-blind, placebo-controlled food challenge in subjects with high levels of IgE antibodies for egg white to compare the allergenicities of heated and ovomucoid-depleted egg white, freeze-dried egg white, and heated egg white. Levels of IgE antibodies for egg white, ovomucoid, ovalbumin, ovotransferrin, and lysozyme were measured in serum by RAST.. Twenty-one of 38 subjects with positive challenge responses to freeze-dried egg white had negative challenge responses to heated egg white, whereas 16 of 17 subjects (94.1%) with positive responses to heated egg white did not respond to the heated and ovomucoid-depleted egg white challenge. The subjects with positive challenge responses to freeze-dried egg white tended to have higher IgE antibody values to ovomucoid than those with negative responses. IgE antibody levels to ovomucoid were significantly higher in subjects with positive responses to a challenge with heated egg white than in those with no response. There were no significant differences in the levels of IgE antibodies to the other proteins, except ovomucoid, in the negative-response and positive-response groups in challenge tests with freeze-dried and heated egg white.. The heated and ovomucoid-depleted egg white preparation was less allergenic than heated or freeze-dried preparations. Ovomucoid has a more important role in the pathogenesis of allergic reactions to egg white than other proteins in egg white.

Topics: Allergens; Antibodies, Anti-Idiotypic; Antibody Specificity; Child; Child, Preschool; Conalbumin; Double-Blind Method; Egg White; Electrophoresis, Polyacrylamide Gel; Female; Food Hypersensitivity; Hot Temperature; Humans; Infant; Male; Muramidase; Ovalbumin; Ovomucin; Placebos; Sodium Dodecyl Sulfate

Various processing aids and fining agents are used in winemaking to help improve sensory characteristics. Some of these materials may contain or be derived from allergenic foods, such as eggs. In order to ensure food safety and that products meet regulatory compliance, it is essential to have robust and effective analytical methods to verify the removal of allergenic proteins following their use. Current methods include ELISA and MS methods, which can target either whole foods or individual proteins, and provide either quantitative data or qualitative confirmation of proteins. MS methods offer the potential to test for multiple proteins within a single assay to improve cost and efficiency, whereas ELISA methods typically analyze for a single protein per assay.. This study focuses on the development of a LC-tandem MS (MS/MS) quantitative method for lysozyme in white wine and compares performance across two laboratories utilizing two different instrument platforms.. Lysozyme target peptides were selected by conducting bottom-up discovery proteomics. Candidate targets were evaluated using parallel reaction monitoring (PRM) or selected reaction monitoring (SRM) LC-MS/MS, depending on the instrument in each laboratory. Quantification of lysozyme was conducted using internal, stable isotope-labeled synthetic peptide standards.. Three of eight candidate target peptides showed performance suitable for the final quantitative method. White wine spiked with 0.1 and 0.5 ppm lysozyme demonstrated quantitative recovery of 70-120%. While the PRM method delivered better repeatability, the SRM method gave higher quantitative recovery values.. A targeted LC-MS/MS method for quantification of lysozyme in white wine has been developed and deployed on two different MS instrument platforms in two laboratories.. Both SRM and PRM targeted LC-MS/MS methodologies can be used for quantification of lysozyme in white wine. This study is among the first to evaluate an MS method for food allergen quantification in multiple laboratories.

Topics: Chromatography, Liquid; Food Hypersensitivity; Humans; Muramidase; Tandem Mass Spectrometry; Wine

Potential residues of the potent allergen lysozyme used as a microbial stabilizing agent in wine production might pose a serious health thread to susceptible individuals. Therefore, EU legislation requires the labeling of the allergenic agent, if it is present in the final product. To allow for product testing, an indirect ELISA method to be specifically used in wine analysis was developed and validated. Furthermore, trial wines treated with defined amounts of lysozyme were subjected to an array of different filtration and other enological processing regimes in order to evaluate their potential to deplete the allergen content of the wines. By these means, processing methods ought to be identified that can be integrated in a good manufacturing practice guideline to enable wine producers to utilize lysozyme in their cellars and still provide wines free of allergenic residues. However, among the enological procedures under scrutiny, only bentonite fining proved to be capable of significantly reducing the allergenic residues.

Topics: Allergens; Bentonite; Egg Proteins, Dietary; Enzyme-Linked Immunosorbent Assay; European Union; Fermentation; Filtration; Food Additives; Food Contamination; Food Handling; Food Hypersensitivity; Food Inspection; Germany; Humans; Microbial Viability; Muramidase; Saccharomyces cerevisiae; Wine

Oral immunotherapy for food allergy has been the focus of a lot of attention recently. The patients have to eat allergenic food instead of eliminating it in this therapy and there is no established standard method yet. To promote clear understanding and improvement of oral immunotherapy, the present study using B10.A mice investigated the effect of multiple oral administration of a model antigen, egg-white lysozyme, on both the antibody response and the anaphylactic reaction induced by subsequent administration of lysozyme. Various doses of egg-white lysozyme (0-100 mg/mouse) were administered to mice intragastrically for 6 d; then additional lysozyme was administered via the intraperitoneal route in all groups. Lysozyme-specific antibody responses were promptly induced by the first oral administration and enhanced by intraperitoneal administration. An anaphylactic reaction was further induced in these sensitized mice by intragastric administration of lysozyme, and the symptoms of shock were compared in order to evaluate the effects of pretreatment. Interestingly, the decrease in rectal temperature which is one of the common anaphylactic symptoms in mice was suppressed in all of the oral pre-administration groups, and the effects were highest in the group that received 20 mg. Consequently, this study using B10.A mice has shown that sensitization can be induced by intragastric administration of lysozyme instead of oral tolerance; however, anaphylactic shock induced by subsequent intragastric administration of lysozyme is suppressed. This mouse model would be useful for assessing the method of oral immunotherapy.

Topics: Administration, Oral; Anaphylaxis; Animals; Dose-Response Relationship, Drug; Egg White; Female; Food Hypersensitivity; Immunotherapy; Injections, Intraperitoneal; Mice; Mice, Inbred Strains; Muramidase

A novel noncovalently bilayer-coated capillary using cationic polymer polybrene (PB) and anionic polymer (sodium 4-styrenesulfonate) (PSS) as coatings was prepared. This PB-PSS coating showed good migration-time reproducibility for proteins and high stability in the range of pH 2-10 and in the presence of 1M NaOH, acetonitrile and methanol. Capillary electrophoresis with PB-PSS coated capillaries was successfully applied to quantitatively investigate the stability of bovine serum albumin, ovomucoid, β-lactoglobulin and lysozyme in simulated gastrointestinal fluids. β-lactoglobulin A and β-lactoglobulin B were both stable in simulated gastric fluid with degradation percentages of 34.3% and 17.2% after 60min of incubation, respectively. Bovine serum albumin, ovomucoid and lysozyme were stable in simulated intestinal fluid with degradation percentages of 17.7%, 23.4% and 22.8% after 60min of incubation, respectively. The superiority of the proposed method over sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and capillary electrophoresis with untreated fused silica capillaries was demonstrated and emphasized.

Topics: Allergens; Animals; Body Fluids; Cattle; Digestion; Electrophoresis, Capillary; Food Hypersensitivity; Gastrointestinal Tract; Hexadimethrine Bromide; Lactoglobulins; Models, Biological; Muramidase; Ovomucin; Polymers; Protein Stability; Proteins; Reproducibility of Results; Serum Albumin, Bovine; Sulfonic Acids

Hidden allergens are a common problem in food safety that has been known for many years. This is why the European Parliament adopted Directive 2003/89/EC amending 2000/13/EC. In addition to specific ingredients, Directive 2003/89/EC also requests the declaration of specific products that were used in the production and could be a risk for allergic individuals. This also includes the declaration of fining agents and lysozyme used in wines. In fact, it could be assumed that fining agents would be almost completely removed during the manufacturing process; however, until now there has been no necessity to analyze wine for these fining agents. By applying enzyme-linked immunosorbent assay (ELISA), residuals of fining agent proteins and the stabilizer lysozyme were investigated in various German wines. The results showed no detectable amounts of fining agents in wines, except for dried egg white and lysozyme, both derived from hen's egg white. For those products, adverse reactions against treated wines could not be excluded.

Topics: Allergens; Egg White; Enzyme-Linked Immunosorbent Assay; Food Handling; Food Hypersensitivity; Food Labeling; Muramidase; Wine

Topics: Cheese; Chromatography, High Pressure Liquid; Egg White; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Food Preservatives; Humans; Muramidase; Reproducibility of Results; Sensitivity and Specificity

To investigate the effect of polysaccharide attachment to proteins on the production of IgG and IgE, the genetic attachment of polysaccharide to lysozymes (G49N and R21T) using the yeast expression system (Saccharomyces cerevisiae AH 22) and the Maillard-type polysaccharide attachment to native lysozyme and soybean P34 protein were attempted. The production of IgG and IgE was investigated by using mice immunized with the protein-polysaccharide conjugates or native proteins. The attachment of polysaccharide to lysozyme using the yeast expression system greatly suppressed the production level of IgG and IgE. The attachment of polysaccharide to native lysozyme and soybean P34 protein using the Maillard-type reaction was also found to be effective in reducing the production level of IgE compared to IgG.

Topics: Electrophoresis, Polyacrylamide Gel; Food Hypersensitivity; Gene Expression; Immunoglobulin E; Immunoglobulin G; Maillard Reaction; Muramidase; Polysaccharides; Saccharomyces cerevisiae; Soybean Proteins

A high-performance and rapid chemiluminescence immunoassay for model food allergen lysozyme, one of the major allergenic components in egg white, using antibody-conjugated bacterial magnetic particles and a fully automated system was developed. This system contains a reaction station, tip rack, and an eight-tip pipettor that is able to attach and detach a strong magnet to the tip surface. The immunoreaction time was shortened to 5 min, and the assay was completed within 20 min. The lower detection limit for lysozyme was 10 ng/mL. This system can be used to perform 24 samples in 60 min within 10% coefficient of variation.

Topics: Allergens; Animals; Chickens; Egg Hypersensitivity; Egg White; Food Hypersensitivity; Humans; Immunoassay; Luminescent Measurements; Magnetics; Models, Immunological; Muramidase

An egg protein, lysozyme, is a still unlabeled additive currently used in cheese preparation. Furthermore, the WHO-FAO committee considers it innocuous. However, 31% of children and 8% of adults with food allergies are allergic to eggs. This work aimed to determine the percentage of patients sensitized to lysozyme from a population of egg-allergic patients. Specific IgE was determined with Cap RAST in 52 patients clinically allergic to egg. Thirty-five percent of egg-allergic patients had antilysozyme IgE. Given this high incidence of lysozyme sensitization, it seems that the presence of lysozyme should be indicated on food labels.

Topics: Administration, Oral; Adolescent; Adult; Antibody Specificity; Child; Child, Preschool; Egg Proteins, Dietary; Egg White; Food Additives; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Lactalbumin; Middle Aged; Muramidase; Prevalence; Radioallergosorbent Test; Skin Tests

Proteins of hen egg whites are common ingredients in food and difficult to eliminate. Allergens of egg white induce allergic symptoms among relatively high numbers of patients suffering from food allergy. B cell epitopes to hen egg white major allergens have been reported. Considering that IgE antibody formation is mostly T cell dependent, the study of T cell epitopes is essential for both T cell dependent and independent IgE response.. Little information on T cell epitopes recognizing food allergens has been reported. T cell responses to hen egg white allergens and two synthetic OA peptides located at amino acid residues No. 105-122 and 323-339 were investigated.. Peripheral blood mononuclear cells from hen egg allergic patients were investigated. Various allergens of hen egg white were used for stimulation. Primary proliferation responses were detected followed by the generation of long-term cultures which were examined for their specificity, phenotype, cytokine profile and IgE production. The allergen specific T cell lines were mapped using a panel of 13 synthetic peptides of ovalbumin.. Human T cells recognizing ovomucoid, lysozyme and ovalbumin epitope 105-122 are reported for the first time. The cell lines were enriched CD4+/CD8+ T cells (CD2+ > 95%). Ovomucoid and ovalbumin induced IgE synthesis by a small fraction of B cells (1%) present in the ovalbumin and ovomucoid specific T cell lines.. Human T cells recognized several egg white allergens and epitopes within the ovalbumin molecule. Specific IgE was produced in cultures stimulated with ovalbumin and ovomucoid. OA peptides 105-122 and 323-339 have no affinity to the specific IgE of the two patients; an observation which could be of particular interest regarding the mechanisms of peptide-based immunotherapy.

Topics: Cell Line; Cells, Cultured; Cytokines; Eggs; Epitopes; Food Hypersensitivity; Humans; Immunoglobulin E; Immunophenotyping; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Muramidase; Ovalbumin; Ovomucin; Peptide Fragments; Radioallergosorbent Test; T-Lymphocytes

IgE is considered to be involved in immediate hypersensitive reactions (IHR) following egg ingestion. IgE antibody levels to egg-white (EW) antigens in the IHR-positive group (n = 19, mean age +/- SD = 5.2 +/- 4.5 yr) were higher than those in the IHR-negative group (n = 13, mean of age +/- SD = 3.6 +/- 2.2 yr). However, even in the IHR-negative group, some patients showed high IgE to EW. RAST inhibition tests with heat-treated (100 degrees C, 5, 10, and 30 min) egg-white antigens were performed on 13 serum samples from subjects with IHR and 9 serum samples from subjects without IHR. Heat treatment decreased the IgE-binding activity of egg white and it was speculated that IgE from IHR-negative subjects bound to relatively heat-unstable sites of egg-white antigens. Furthermore, we selected IHR-negative subjects (n = 8, mean of age +/- SD = 3.0 +/- 1.7 yr) with higher IgE antibody levels than the lowest limit of IgE to EW of the IHR-positive group and compared IgE to ovomucoid (OM), ovalbumin (OA), conalbumin (CA), and lysozyme (Ly) between these IHR-negative and positive groups. IgE-binding activities to egg-white components, including OA, CA, and Ly but not OM, were significantly decreased with heat treatment. The IHR-negative group showed significantly lower IgE to OM (untreated, 5, 10, 30 min treatment) and 5 min treated OA alone than the IHR-positive group, while no difference was found in IgE to other components between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Antigen-Antibody Reactions; Antigens; Child; Child, Preschool; Conalbumin; Egg White; Female; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Infant; Male; Muramidase; Ovalbumin; Radioallergosorbent Test

Hen's egg white lysozyme (HEL) is one of the minor allergen in hen's egg white. HEL is commonly used to treat disease of respiratory tract, because it have the effect to dissolve mucopolysaccharide and anti-inflammatory action. We examined specific IgE antibody titers (IgE-HEL) in patients with egg allergy and allergic patients to other antigen than egg. Results indicated that 16.37 +/- 29.56 (PRU/ml) (mean +/- SD) of IgE-HEL was found in 30 out of the 39 allergic patients to egg, and 23 (66.7%) out of the 39 patients studied showed RAST scores of more than 2. On the other hand, 1.08 +/- 0.92 (PRU/ml) of IgE-HEL in 12 out of the 44 allergic patients to other antigen than egg, and 5 (11.4%) out of the 44 patients studied showed RAST scores of more than 2. Moreover, we treated a patient who developed anaphylaxis after taking HEL. 1.0 (PRU/ml) of HEL-IgE was found in this patient. These results suggest that we should be careful in treating allergic patients with HEL.

Topics: Adolescent; Age Factors; Anaphylaxis; Antibody Specificity; Child; Child, Preschool; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Male; Muramidase; Peptide Fragments; Radioallergosorbent Test

The radioallergosorbent test (RAST) and RAST inhibition test were used to examine cross-allergenicity amongst the major hen's egg-white and egg-yolk proteins. Using ovalbumin as a reference allergen to compare cross-reactivity, it was apparent that the proteins conalbumin, ovomucoid and lysozyme substantially inhibited binding to ovalbumin discs of IgE in the sera of patients clinically hypersensitive to egg. The converse situation with conalbumin, ovomucoid and lysozyme on the discs and ovalbumin as the inhibitor also resulted in significantly decreased levels of IgE binding to the proteins on the discs. It was also demonstrated that cross-reactions occurred between ovalbumin and the yolk protein, apovitellenin I. Cross-reaction was also observed surprisingly when egg lysozyme was on the disc and the milk protein allergen alpha-lactalbumin was used as the inhibitor. The demonstration of cross-reaction between all of these proteins may signify that there are a number of common allergenic determinants on these egg proteins, thus providing a molecular basis for the phenomenon of cross-reactivity.

Topics: Adolescent; Adult; Apoproteins; Child; Child, Preschool; Cross Reactions; Egg Proteins; Egg Proteins, Dietary; Food Hypersensitivity; Humans; Lactalbumin; Muramidase; Ovalbumin

Topics: Adolescent; Antibody Specificity; Child; Child, Preschool; Egg White; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Male; Muramidase; Radioimmunoassay

The radioallergosorbent test (RAST) was used to compare the IgE binding of egg white and yolk, and allergenic proteins were detected by immunoelectrotransfer ('Western blotting'). The main allergens were found in egg white, but for a large proportion of the egg-sensitive patients, yolk contained specific IgE-binding constituents. For blood sera from 36 patients, there was a positive correlation between the results of RAST for egg white and for yolk. Lysozyme was found to be an allergen for some patients. The effect of heating on the allergenicity of egg white was examined and the allergenicity of hen egg white was compared with that of a duck egg. The allergens in yolk were associated with each of the three yolk fractions, and several of the proteins in the low-density lipoprotein fraction bound IgE.

Topics: Adolescent; Adult; Allergens; Animals; Chickens; Child; Child, Preschool; Egg Proteins; Egg White; Egg Yolk; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Male; Muramidase; Radioallergosorbent Test

Topics: Adolescent; Adult; Aged; Animals; Bacterial Infections; Child; Child, Preschool; Dogs; Ear, Middle; Eustachian Tube; Food Hypersensitivity; Gases; Humans; Hypersensitivity; Immunoglobulin E; Infant; Lymph; Middle Aged; Mucus; Muramidase; Otitis Media; Otitis Media with Effusion; Perilymph

Topics: Allergens; Animals; Chickens; Egg White; Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Female; Food Hypersensitivity; Humans; Immune Sera; Immunoelectrophoresis, Two-Dimensional; Isoelectric Focusing; Muramidase; Ovalbumin; Ovomucin; Rabbits; Radioallergosorbent Test

Allergens in hen's egg white were studied in crossed radio-immunoelectrophoresis (CRIE). Specific IgE-antibodies against different proteins in the egg white were examined in sera from 70 atopic patients with clinical hypersensitivity, and RAST greater than or equal to 2 to egg white. Ovalbumin, ovomucoid and an unidentified protein, antigen 22, were classified as major allergens. Specific IgE-antibodies against 10 more proteins in hen's egg white were detected. IgE-antibodies against lysozyme could not be detected.

Topics: Adolescent; Allergens; Animals; Chickens; Child; Child, Preschool; Egg White; Female; Food Hypersensitivity; Humans; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Infant; Muramidase; Ovalbumin; Ovomucin; Rabbits; Radioallergosorbent Test; Skin Tests

Topics: Animals; Antibodies, Viral; Arteritis; Colitis, Ulcerative; Crohn Disease; Cytomegalovirus; Food Hypersensitivity; Humans; Immunity, Cellular; Lupus Erythematosus, Systemic; Mice; Milk; Muramidase; Rabbits; RNA Viruses