IgE antibodies to wheat flour components Studies with sera from subjects with bakers' asthma or coeliac condition

Sera from two subjects with bakers' asthma and six patients with coeliac condition were examined for the presence of IgE antibodies with specificities for wheat flour components. Sera were studied using the radioallergosorbent test (RAST) together with whole flour and thirteen purified and partially purified flour fractions. IgE antibodies to a number of flour components were demonstrated in the allergic bakers' sera, but the strongest reactivities were observed with wheat albumins and globulins. A more detailed examination of the flour water-soluble proteins using RAST inhibition methods demonstrated that albumins were more reactive with the allergic sera than the globulins. Apart from the results with the water-soluble proteins, the two sera showed a different pattern of reactivity with the other flour preparations. No IgE antibodies to whole flour or any of the flour components, including A gliadin, were found in the coeliac sera. Failure to detect wheat gluten- or gliadin-specific IgE antibodies indicates that IgE-mediated reactions are not important in the pathogenesis of coefiac condition. Levels of total IgE in the sera from the coeliac subjects were elevated and, with two of the sera, some success was achieved in identifying the allergens responsible for the elevation. We propose that elevation of serum IgE may frequently occur in coeliac condition and may arise due to an increased uptake of antigens via the damaged intestinal mucosa.     

Immunoglobulin E specific to wheat and rye flour proteins

We have used the radioallergosorbent test (RAST) to determine IgE antibodies specific to wheat flour proteins in the sera of seven groups of patients. In some cases rye-specific IgE was also determined. Wheat and rye RAST scores showed a good correlation, presumably due to cross-reactions. Among bakers with asthma, positive scores, 0.5–3, occurred with a prevalence of 43%, and among children with eczema, scores in the range 0.5–4 were found with a prevalence of 54%. A score of 0.5 was a marginal value which was also occasionally encountered with sera from patient groups with no history of immediate hypersensitivity to wheat or rye. These groups included adults and children with allergic rhinitis and asthma, children from the general population and children with coeliac disease. The RAST appeared useful in the diagnosis of allergy to inhaled flour dust among bakers. Among children with eczema, positive wheat and rye RAST results were a common finding, which only occasionally could be linked to strong and unequivocal reactions to the foods in question. Both in bakers and children with eczema, wheat and rye RAST results showed good agreement with intracutaneous skin test results.     

Immunoglobulin E antibodies against Staphylococcus aureus cell walls in the sera of patients with hyperimmunoglobulinemia E and recurrent staphylococcal infection.

The specificity of antistaphylococcal antibodies of the IgE class in five patients with hyperimmunoglobulinemia E and recurrent staphylococcal infection has been investigated. Purified cell walls were prepared from various staphylococcal strains, and serum immunoglobulin E binding was measured by using a solid-phase radioimmunoassay. Immunoglobulin E binding occurred only with cell walls from Staphylococcus aureus strains, including walls from a teichoic acid-deficient mutant. Immunoglobulin E did not bind to cell wall preparations from the coagulase-negative species S. capitis, S. sciuri subsp. lentus, S. simulans, S. xylosus, staphylococcal strains RB-11 and Armour, and from a group A streptococcus strain CS44. Since the glycan backbone and the tetrapeptide (pentapeptide) subunit of the peptidoglycan of all staphylococcal strains tested are believed to be identical, it is suggested that IgE binding is related to either the peptidoglycan interpeptide bridge or an unknown antigenic structure within the cell wall of S. aureus. The pathophysiological significance of antistaphylococcal immunoglobulin E antibodies in the disorder studied is at present unknown. The formation of immunoglobulin E antibodies to S. aureus cell wall components may be a manifestation of an aberrant immunological response to S. aureus related to the undue susceptibility to staphylococcal infections in these patients.     

Immunoglobulin E reactivity to latex antigens in the sera of patients from Finland and the United States

Background: Patients with latex sensitivity and latex antigens from the United States and Finland, two countries where allergic reactions to latex have been widely reported, were evaluated to determine the spectrum of immune responses.Methods: Sera from 27 patients from Finland and 18 from the United States with latex allergy and control sera from nonsensitive individuals were studied for latex-specific IgE antibodies. Four antigen preparations were used: two extracted from gloves and one each extracted from rubber tree sap from Malaysia and India. All 45 patients had skin prick test results that were positive to latex antigens, and all sera were evaluated by enzyme-linked immunosorbent assay (ELISA) with the various antigens.Results: There were considerable differences in the reactivity of patient sera with the different antigens. Only 50% of the sera from patients with latex allergy from Finland demonstrated significant levels of IgE to latex as determined by enzyme-linked immunosorbent assay. These patients showed more reactivity with rubber tree sap antigens than with glove antigens. However, 72% of the patients from the United States demonstrated antibodies to latex, and no marked differences were noted between the antigen extracts.Conclusions: The results indicate that reagents such as rubber tree sap, which contain multiple clinically significant antigenic components, should be included in evaluation of latex allergy and that differences in patient populations may result in serologic variances.     

Identification of crossreacting wheat, rye, barley and soya flour allergens using sera from individuals with wheat-induced asthma

We have conducted radio allergosorbent test (RAST), competitive RAST inhibition, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting using sera from patients with wheat-induced asthma to investigate the immunological relationship between wheat, rye, barley and soya, and to identify common proteins between these flours. RAST showed strong associations between the levels of specific IgE to wheat flour and those of rye and barley flour. Competitive RAST inhibition showed that wheat, rye, barley and soya flours contained crossreacting proteins, in decreasing concentrations. Wheat, rye and barley flours had similar protein profiles on gel electrophoresis. Soya flour contained a number of high molecular weight proteins not present in the other cereals. Western blotting using sera from 21 wheat flour hypersensitive individuals identified a large number of allergens in the different flours. Proteins of 69, 33, 26, 21 and 12 kDa were identified as major wheat flour allergens. Rye flour proteins of 21 and 12 kDa, and barley flour proteins of 69, 52 and 10 kDa were the major allergens identified by serum from wheat hypersensitive individuals. The major common protein of soya and wheat flour had a molecular weight of 21 kDa. The majority of crossreacting allergens identified between the different flours have molecular weights similar to those of known flour enzymes or enzyme inhibitors.     

Immunoglobulin class of antibodies to cow's milk casein in infant sera and evidence of low molecular weight IgM antibodies

Use of the red cell linked antigen—antiglobulin reaction with specific antiglobulin sera allows the recognition of the immunoglobulin class of the reacting antibody.In the youngest infants IgG was the only immunoglobulin reacting with casein. With the particular sera studied the antibody measured was probably maternal. In older infants IgA and IgM antibodies were also detected but they were usually present at lower levels than IgG antibodies.In serum from a boy with Aldrich''s syndrome the IgA level was very high and IgA antibody to casein (titre 128,000) was higher than the IgG antibody.In selected sera containing IgM antibody to casein, evidence is produced of a low molecular weight form of μ-specific antibody. In Sephadex G-200 filtration it was located in the `7S'' fraction containing IgG antibody.     

Anti-thyroid peroxidase antibodies in sera from healthy subjects and from patients with chronic thyroiditis: differences in the ability to inhibit thyroid peroxidase activities.

A significant percentage (6.4%) of healthy subjects was found to contain anti-thyroid peroxidase (TPO) antibodies in their sera. However, in contrast with IgG from sera of patients with chronic thyroiditis, IgG from sera of healthy subjects did not inhibit TPO activities both in guaiacol and iodide assays. In addition, anti-TPO antibodies from healthy subjects did not block the inhibition of enzyme activities by anti-TPO antibodies from patients. These findings suggest that anti-TPO antibodies from healthy subjects do not bind to the epitopes relating to substrate-combining sites of TPO. Thus, the specificities of anti-TPO antibodies in healthy subjects may differ from those in cases of chronic thyroiditis.     

Immunoglobulin E and immunoglobulin G4 antibodies to cow''s milk in children with cow''s milk allergy

The presence of cow's milk specific antibodies of immunoglobulin E and G4 classes were studied in 47 children with a positive clinical history of cow's milk allergy. The children were challenged with cow's milk orally. The clinical diagnosis was verified by immediate reactions in 25 patients while 22 had late reactions or were provocation test negative in spite of the clinical history. There was no relation between levels of cow's milk specific IgG4 antibodies and provocation test result, i.e. neither with immediate or late reactions. Total IgF. Was elevated above + 1 SD for age in 31 41 tested patients. Of these, 29 had immediate type reactions to cow's milk wheat flour and/or egg white, while only two of 10 children with IgE of less than +1 SD had a demonstrable allergy to any of these foods. The sensitivity was 80%. Specific IgE antibodies to cow's milk were demonstrated in 11 of 14 children with immediate reactions and in three of 15 who were provocation test negative or had only late reactions. This means a sensitivity of 79% and a specificity of 80%. At least had only late reactions. This means a sensitivity of 79% and a specificity of 80%. At least one of the four patients with specific IgE but negative provocation test result had earlier shown an immediate reaction when challenged with cow's milk indicating that the specific IgE antibodies were not truly "false" positive reactions but a consequence of previous allergy. Our results confirm an association between elevated total serum IgE and food allergy and an association between positive RAST to cow's milk and positive provocations in young children. We did not find any evidence for specific IgE-1 antibodies playing a role in these patients.     

Wheat gliadin fractions and other cereal antigens reactive with antibodies in the sera of coeliac patients.

The mixed reverse solid phase passive antiglobulin haemadsorption test (MRSPAH) and the enzyme linked immunosorbant assay (ELISA) were found equally sensitive and fitted for the measurement of serum IgG antibodies against alcohol soluble gliadins. Using the ELISA method, three coeliac sera with elevated antibody titres against gliadins and two control sera with low titres were tested for IgG antibodies against the main groups of wheat proteins (acetic acid soluble glutenins, salt soluble albumins and globulins and alcohol soluble gliadins), eight fractions of gliadin and the alcohol soluble proteins of barley, rye, oat, maize and rice. As rice contained little alcohol soluble protein, a test against acid soluble rice proteins was included. In all three patient sera, titres higher than or equal to that for crude gliadin were found for wheat glutenin and for gliadin fractions seven and eight, both containing alpha gliadins. Similar high titres were found when these coeliac sera were tested against rye, barley and oat prolamines. Maize prolamines gave only low titres and no antibodies could be detected against rice proteins, in line with the tolerance of these latter two cereals by patients with coeliac disease. It would appear that sera from coeliac patients react with more than one antigenic fraction of protein in wheat and other cereals. Also sera from two normal persons appeared to have the same spectrum of reactivity against these cereal proteins as did the three sera from coeliac patients. The titres in normal sera were however much lower.     

Immunoglobulin E-mediated asthma and hypersensitivity pneumonitis with precipitating anti-hapten antibodies due to diphenylmethane diisocyanate (MDI) exposure

Two workers are presented who were exposed to diphenylmethane diisocyanate (MDI) while coating pipes with a polyurethane foam. After a latent period of exposure, worker A developed immediate-onset asthma and worker B developed a clinical picture of hypersensitivity pneumonitis for which he was hospitalized. The antibody response of these workers to a conjugate of MDI with human serum albumin (MDI-HSA) was measured by gel precipitation, total antibody binding of 125I MDI-HSA, and specific IgG and IgE antibody by polystyrene-tube radioimmunoassay (PTRAI). Worker B had precipitating antibody to MDI-HSA by double immunodiffusion in gel. Both workers had high levels of IgG antibody specific for MDI-HSA which had some cross-reactivity with a conjugate or toluene diisocyanate and HSA. Total serum antibody binding of 125I MDI-HSA was 15 microgram/ml in worker A and 900 microgram/ml in worker B. Both workers had serum IgE antibody specific for MDI-HSA as measured by two PTRIA techniques. These results indicate that a marked immunologic response to MDI is possible in exposed workers and that hypersensitivity pneumonitis can occur subsequent to the inhalation of a low-molecular-weight chemical in the industrail setting.     

Detection of antibodies to human coronaviruses 229E and OC43 in the sera of multiple sclerosis patients and normal subjects.

Sera collected from 90 multiple sclerosis patients and 148 age-matched normal subjects were examined for the presence of antibodies against human coronaviruses (HCV) 229E and OC43 by enzyme immunoassay (EIA). The results demonstrated no significant difference between the MS patients and the normal subjects in their antibody titer to HCV 229E and HCV OC43. Further analysis of these 238 sera indicated that a stronger EIA reaction was generally observed against HCV OC43 (mean EIA value at an optical density of 492 nm = 0.896) than against HCV 229E (mean EIA value at an optical density of 492 nm = 0.346).     

Catfish antibodies to blood group substances: III. Further studies with sera from fish injected with human group O secretor fluids

An antiserum prepared in the freshwater catfish Tandanus tandanus by the injection of O secretor seminal plasma was fractionated into anti-H reagents showing different specificities by absorption with A₁B erythrocytes and by absorption and elution from A₁B cells. Although purified human and hog H blood group substances inhibited the haemagglutination of O erythrocytes by both the eluate from A₁B cells and the serum remaining after absorption with A₁B cells, all of the simple sugars tested, except 2′-fucosyl-lactose, failed to inhibit either sample. The H-substances inhibited the A₁B-eluate at dilutions which were significantly higher than those required to inhibit the A₁B-absorbed serum. Inconsistent with this result was the finding that 2′-fucosyl-lactose, a trisaccharide with a structure similar to the terminal H-active groupings on the type 2 chains of the ABH macromolecules, was a more active inhibitor of the absorbed than of the eluted serum. Seventeen different samples of O secretor saliva either failed to inhibit the A₁B-absorbed serum, or produced inhibition at very low dilution. These same saliva samples inhibited the A₁B-eluate in high dilution.     

Immunoglobulin E antibodies that crossreact with vegetable foods,pollen, and Hymenoptera venom

IgE in some human sera reacted with an antigen present in a large number of unrelated foods: potato, spinach, wheat, buckwheat, peanut, honey, and others. The antigen, which was periodate-sensitive and heat-stable, was also found in pollen. Even more surprisingly, these antibodies often reacted in vitro with bee and vespid venom and were sometimes apparently induced by Hymenoptera stings.     

The effects of inhaled leukotriene E4 on the airway responsiveness to histamine in subjects with asthma and normal subjects

Eight subjects with asthma inhaled on separate occasions leukotriene E4 (LTE4) (6.1 nmol, geometric mean), methacholine, and diluent, which produced an average 41.0%, 37.0%, and 3.3% decrease in specific airway conductance (SGaw), respectively. When the SGaw had recovered to baseline levels at 60 minutes after challenge, the provocative dose of inhaled histamine that produced a 35% decrease in SGaw (PD35) was determined. The histamine PD35 observed after inhalation of LTE4 was 0.46 mumol, and this was significantly less than the histamine PD35 observed after inhalation of methacholine (0.88 mumol; p less than 10(-4) and diluent (0.97 mumol; p less than 10(-5). Histamine responsiveness was also enhanced by a fiftyfold lower dose of LTE4 (p = 0.005), and the enhancement was less than that elicited by the higher dose of LTE4 in the same individuals (p = 0.02). The changes in histamine PD35 during a 1-week period after LTE4 and methacholine challenges were compared in four subjects with asthma. There was a time-dependent enhancement in histamine responsiveness that reached a maximal of 3.5-fold at 7 hours after LTE4. The enhancement had disappeared by 1 week. Similar changes were not observed after methacholine challenge, which elicited the same degree of bronchoconstriction as LTE4. Inhalation of LTE4 in five normal subjects that produced a mean 37.6% decrease in SGaw did not change histamine responsiveness for up to 7 hours. These findings suggest that LTE4 may play a role in the perpetuation of nonspecific airway hyperresponsiveness in bronchial asthma.     

Immunoglobulin G antibodies against indoor dampness-related microbes and adult-onset asthma: a population-based incident case-control study

Immunoglobulin G (IgG) antibodies against microbes related to indoor dampness problems have been used as potential biomarkers of fungal exposure in clinical investigations. There is limited information on their relation to asthma. We conducted a population-based incident case-control study to assess the risk of asthma in relation to specific IgG antibodies to eight dampness-related microbes: Aspergillus fumigatus, A. versicolor, Cladosporium cladosporioides, Fusarium oxysporum, Sporobolomyces salmonicolor, Stachybotrys chartarum, Streptomyces albus and Trichoderma citrinoviride. We recruited systematically all new cases of asthma during a 2.5-year study period and randomly selected controls from a source population of adults 21-63 years of age living in the Pirkanmaa Hospital District, South Finland. The clinically diagnosed case series consisted of 521 adults with newly diagnosed asthma and the control series of 932 controls selected randomly from the source population. IgG antibodies were analysed with ELISA. An increased risk of developing asthma in adulthood was significantly related to IgG antibodies to T. citrinoviride, but not to the other moulds. There was no evidence of a dose-response relation between the IgG antibody level and the risk of asthma. T. citrinoviride may play a role in the aetiology of adult-onset asthma or serve as an indicator of other causal factors.     

Crossreactivity of IgE antibodies from sera of subjects allergic to both ryegrass pollen and wheat endosperm proteins: evidence for common allergenic determinants

Positive RAST (greater than 5% radioactive uptakes) to wheat endosperm proteins were found in approximately one-quarter of subjects who had both a positive skin prick and RAST (greater than 10% radioactive uptake) to ryegrass pollen proteins. Immunoblotting of proteins electrophoretically transferred to nitrocellulose membrane after SDS-PAGE of ryegrass pollen and wheat endosperm proteins confirmed the crossreactive properties of the sera identified by RAST testing. Immunoadsorption of serum IgE onto nitrocellulose membrane, to which ryegrass pollen or wheat endosperm proteins had been adsorbed, removed IgE from crossreactive sera reactive to both ryegrass pollen and wheat endosperm proteins. Elution of the adsorbed IgE from the nitrocellulose membrane after immunoadsorption and probing blotted strips of both ryegrass pollen and wheat endosperm proteins supported the results obtained from the immunoadsorption experiments. This data provides evidence that the crossreactivity of IgE antibodies in sera reacting with both ryegrass pollen and wheat endosperm proteins involves common or related determinants and has implications for the clinical management of these allergic subjects.     

Immune complexes and antibodies to BCG in sera from patients with mycobacterial infections.

We have examined the prevalence of circulating immune complexes in sera from patients with mycobacterial infections. Sera from 68 percent of patients with active M. tuberculosis and 58 percent of patients with M. intracellulare infections had significantly elevated Clq binding activity (Clq-BA). In general there was a fall in Clq-BA with treatment. Only 15 percent of M. tuberculosis patient with a bacteriological cure, 22 percent of non-tubercular patients with chronic obstructive pulmonary disease, and 3 percent of normal individuals had an elevated Clq-BA. Antibodies to a BCG-derived antigen were demonstrated in most of the individuals studied in all groups but, significantly elevated levels were seen only in patients with mycobacterial infections. In certain patients there appeared to be an inverse relationship between Clq-BA and BCG binding, suggesting that perhaps BCG-related antigens participated in the immune complexes found. Other possible antigen-antibody complexes are discussed.