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Determination of IgE antibodies to the benzyl penicilloyl determinant. A comparison between poly-L-lysine and human serum albumin as carriers.

The influence of two different carriers, poly-L-lysine (PLL) and human serum albumin (HSA) in the binding of specific IgE antibodies to the benzyl penicilloyl hapten (BPO) was determined in a solid-phase assay. Serum samples from patients with a history of immediate reaction to penicillin and which had shown the presence of IgE antibodies to BPO were used. Benzyl penicilloyl derivatized cellulose discs were prepared using PLL of different molecular weight and HSA as carriers. These were treated with different molar ratios of benzyl penicillin. These reagents were compared for uptake of BPO-specific IgE using a pool of sera in a radioallergosorbent test (RAST) type assay. Two PLL systems and two HSA systems were finally compared using 26 individual sera. RAST values were compared by Kruskal-Wallis and Wilcoxon tests. The relationships between the four different assays were evaluated by determining Pearson correlation coefficients and the concordance by determining intraclass correlation coefficients (ICC). Analysis of means by the Wilcoxon test revealed significant differences (P less than 0.01) only when the different carrier assays were compared. The correlation coefficients between all the assays were significant (P less than 0.0001), but the ICC was low when the different carrier assays were compared. These results indicate that the nature of the carriers studied (PLL and HSA) influences the capacity for binding IgE antibodies in the RAST procedure. The differences observed indicate that one conjugate cannot be substituted for the other in the determination of IgE antibodies to BPO and that BPO-PLL is preferable.

Antibodies↗

Severe hypersensitivity reaction during hemodialysis.

BACKGROUND: Hemodialysis-associated hypersensitivity reactions are well documented in the literature. Ethylene oxide sensitization and activation of complement are important factors involved during such reactions. The majority of severe hypersensitivity reactions in dialysis patients, however, is due to sensitization to ethylene oxide. METHODS: We report a patient admitted to the hospital with worsening of his renal function. He subsequently required three hemodialysis treatments, all of which resulted in severe hypersensitivity reactions requiring endotracheal intubation. The initial hypersensitivity episode was thought to be due to complement activation to the cupramonium-rayon membrane dialyzer. Despite changing to a polyacrylonitrile membrane, which does not activate complement, a second hypersensitivity reaction developed. Suspecting ethylene oxide hypersensitivity, the third hemodialysis session incorporated a biocompatible dialyzer that was sterilized with gamma irradiation, not ethylene oxide. Yet again, an anaphylactoid reaction resulted. It was postulated that residual ethylene oxide in the tubing might have triggered this last attack. RESULTS: Despite a negative RAST (radioallergosorbent test) to ethylene oxide, the strong history surrounding each of the hypersensitivity episodes and high index of suspicion pointed to ethylene oxide hypersensitivity as the etiologic factor. To this end, the patient fared much better when peritoneal dialysis was initiated. The patient subsequently died from other complications of his illness. CONCLUSIONS: This case report demonstrates both the complex nature involving a hypersensitivity reaction to hemodialysis and the life-threatening severity of such a reaction. Replacing ethylene oxide with steam or gamma radiation to sterilize dialyzers and thoroughly rinsing new dialyzers and tubing with normal saline may help circumvent this problem.

Acrylonitrile↗

An investigation of the antigens of Ascaris lumbricoides using a radioimmunoassay and sera of naturally infected humans.

Of 5 humans from an Ascaris lumbricoides(var. suum)-endemic area who were positive for Ascaris infection (past or current) when examined by a radioallergosorbent test (RAST) for serum immunoglobulin E, only 1 contained detectable levels of Ascaris specific serum IgG antibodies. These antibodies were not detectable when the radioimmunoassay was replaced with a precipitin assay. There was a wide ringe of molecular weights and isoelectric points of antigens in Ascaris body fluid (ABF) which reacted with antibodies in this infected human. However, unlike the IgE-binding ABF antigens, the IgG-binding ABF antigens were particularly evident in one area of molecular weight and isoelectric point. This human also had IgA and IgM to Ascaris antigens whereas the others had barely detectable amounts.

Animals↗

Food allergy to wheat: identification of immunogloglin E and immunoglobulin G-binding proteins with sequential extracts and purified proteins from wheat flour.

BACKGROUND: Cereal-associated allergy is particularly considered a serious problem, because cereals are essential in our daily diet. Wheat proteins are classified into albumins, globulins and prolamins (insoluble gliadins and glutenins). OBJECTIVES: Our objectives were to study the involvement in food allergy to wheat of these different protein types by using purified fractions and to identify those binding IgE and IgG antibodies. METHODS: Sera were obtained from 28 patients with food allergy to wheat. Albumins/globulins, gliadins and glutenins were obtained by sequential extraction based on differential solubility; alpha-, beta-, gamma- and omega-gliadins and low molecular weight (LMW) and high molecular weight (HMW) glutenin subunits were purified by chromatography. IgE binding to these extracts and fractions were analysed by radioallergosorbent test (RAST), and immunoblotting; IgG binding was detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: In RAST, 60% of sera were shown to have specific IgE antibodies against alpha-, beta-gliadins and LMW glutenin subunits, 55% to gamma-gliadins, 48% to omega-gliadins and 26% to HMW glutenins. Immunoblotting analysis confirmed results obtained in RAST concerning LMW and HMW glutenin subunits and showed that 67% of patients have IgE antibodies to the albumin/globulin fraction. CONCLUSION: Results obtained in the different tests showed common features and in agreement with other studies indicated the presence of numerous allergens in food allergy to wheat; alpha-, beta-, gamma- and omega-gliadins, LMW glutenin subunits and some water/salt-soluble proteins appeared as major IgE binding allergens, whereas HMW glutenins were only minor allergens. The same type of antigenic profile against gliadins and glutenins was observed with IgG antibodies. Important sequence or structural homologies between the various gliadins and LMW glutenin subunits could certainly explain similarity of IgE binding to these proteins.

Adult↗

Effect of elimination diets on food-specific IgE antibodies and lymphocyte proliferative responses to food antigens in atopic dermatitis patients exhibiting sensitivity to food allergens.

Peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis (AD) selected as being sensitive to hen's egg or cow's milk responded to food antigens, ovalbumin, or bovine serum albumin, with significantly enhanced DNA synthesis compared with the DNA synthesis in PBMCs from nonatopic control subjects and food-sensitive patients with immediate symptoms. Patients were treated with elimination diets. Symptoms of AD had been in remission during elimination diets. The levels of specific IgE antibodies to hen's egg or cow's milk decreased during elimination diets in patients with positive radioallergosorbent test (RAST). In patients with negative RAST, specific IgE antibodies remained negative during elimination diets. The proliferative responses of PBMCs to food antigens also decreased during elimination diets in patients with proliferative responses before elimination diets. Taken together, specific IgE antibodies to food antigens are useful indexes of the effect of elimination diets in food-sensitive patients with AD and positive RAST, and proliferative responses of PBMCs to food antigens are useful indexes of the effect of elimination diets in food-sensitive patients with AD and proliferative responses of PBMCs.

Adolescent↗

Immunologic and biochemical evaluation of the potency of whole insect body extracts.

Recent studies have indicated that currently available whole body extracts have little potency and are ineffective for diagnosis and treatment of stinging insect allergy. Pure venom is a potent effective allergen but is difficult to obtain in sufficient quantities from all Hymenoptera species. In these studies, an attempt was made to prepare a potent whole body extract. Whole bee body extracts were prepared with different extraction periods and at cold and room temperatures. Potency was examined biochemically by measurements of phospholipase A (PLA) activity and immunologically by PLA and bee venom radioallergosorbent test (RAST) inhibition experiments and gel diffusion studies with the use of rabbit antisera. All extracts prepared in the laboratory had some potency, indicating that it is possible to make a whole body extract containing small quantities of PLA or bee venom. However, the potency of these extracts was minimal as compared with bee venom. Three commercial extracts were almost devoid of detectable immunologic activity. While further attempts may be made to prepare a potent whole body insect extract, these results suggest that it is necessary to obtain venom in relatively pure form for the diagnosis and treatment of stinging insect allergy.

Animals↗

Studies on the physicochemical properties of house dust mite (Dermatophagoides pteronyssinus) allergens involved in reaginic reaction.

The physicochemical nature of the allergenic component of Dermatophagoides pteronyssinus extract was studied by gel filtration, ion exchange chromatography, isoelectrofocusing, and polyacrylamide gel electrophoresis. Fractions were assayed for allergenic activity by the direct radioallergosorbent test (RAST) (dilution method), RAST inhibition (50% inhibition method) and human skin testing. Fractions were further studied by preparative polyacrylamide gel electrophoresis and the band showing a flow rate value of 0.35 was the most potent in allergenic activity. Fractionation by isoelectrofocusing yielded three factions with allergenic activity of pI = 4.8, PI = 7.4 and pI = 10.0 Sephadex G-100 gel filtration revealed that allergenic activity was present in fractions with molecular weight ranging from 17,000-45,000 and discrete ion exchange chromatography evidenced three fractions with very high allergenic activity. In the present study we propose that the allergenic material of D.P is composed of 3 physicochemically different substances whose molecular weight lie in the 17,000 to 45,000 range.

Allergens↗

Total and specific IgE in serum and cerebrospinal fluid of rats and guinea pigs infected with Angiostrongylus cantonensis.

The total and specific IgE response to Angiostrongylus cantonensis infection was evaluated according to host permissiveness. Total IgE levels measured by a double antibody radioimmunoassay (RIA) increased slowly in the permissive host (the rat), reaching a maximum between 4 and 8 weeks after infection. This maximum was earlier but significantly lower in the non-permissive host (the guinea pig). IgE antibodies specific for adult worms or L1 or L3 larvae of A. cantonensis were measured by a radioallergosorbent test (RAST). In the case of adult worms and L1 antigens, specific IgE antibody levels showed large variations in relation to the duration of infection in rats. In contrast to total IgE levels, the specific IgE response to L3 larvae was lower in rats than in guinea pigs in both the serum and cerebrospinal fluid (CSF). These results suggest variations in the total vs specific IgE response according to host permissiveness or non-permissiveness to A. cantonensis infection. These results are discussed in the context of the possible participation of IgE antibodies in immune defence.

Angiostrongylus↗

Intestinal anisakiasis. A case diagnosed by morphologic and immunologic methods.

The diagnosis of the fourth reported case of intestinal anisakiasis in the United States was based on the morphologic characteristics of the worm in histologic sections of resected ileum. Detection of antibodies to the worm by a radioallergosorbent test (RAST), enzyme-linked immunosorbent assay (ELISA), and an immunofluorescent antibody assay (IFA) support the diagnosis. IgE antibodies in patient serum reacted specifically to larval Anisakis antigens but not to larval Ascaris antigens in the RAST. IgG and IgM antibodies to larval Anisakis antigen could be detected by ELISA up to six months after infection. IFA findings demonstrated that patient serum recognized the excretory-secretory products produced by the worm and showed their presence in vivo. These products appear to originate from the excretory pore and dorsal esophageal gland of the larval parasite.

Adult↗

Cross-reactivity between Acacia (wattle) and rye grass pollen allergens. Detection of allergens in Acacia (wattle) pollen.

Immunoglobulin E (IgE), directed against components of Acacia (wattle) pollen, has been detected by radioallergosorbent tests (RAST) in the sera of some children and adults who develop allergic symptoms in the presence of flowering Acacia trees in Australia. All these subjects also had high levels of IgE directed against Lolium perenne (rye grass) pollen. Inhibition by RAST showed that most of the IgE molecules which bound to Acacia pollen components also bound to L. perenne pollen extracts, and to Glycoprotein 1, the major allergen of L. perenne pollen. In these assays, the allergens have been immobilized on polyvinyl chloride microtitre trays: the sensitivity of this approach is compared to that of commercial RAST kits.

Acacia↗

The prevalence of latex allergy in operating theatre staff.

Latex allergy has become increasingly common amongst health care workers. The prevalence of latex allergy in 102 theatre personnel at Princess Alexandra Hospital was determined by the results of a standardized questionnaire and a latex specific IgE radioallergosorbent test (RAST). Volunteers had their forced expiratory volume in 1 second/forced vital capacity (FEV1/FVC) measured at the beginning and end of their working shifts. Only one of 102 volunteers had a positive latex specific IgE RAST and he also experienced both local and systemic symptoms with latex exposure and a deterioration in daily FEV1/FVC. The 14 volunteers using asthma medications had significantly higher incidence of operating theatre symptoms (10/14), local reactions (8/14), and atopy (14/14). There was no clinically significant daily change in FEV1 in the total population or any specific group. The 1% (1/102) prevalence of latex allergy and sensitization in theatre staff demonstrates a much lower incidence than previously reported (12.5%, 15.8%).

Adult↗

A study on antigenic and allergenic changes during storage in three different biological extracts.

The stability of three allergens common in tropical countries was evaluated under different storage conditions. Prosopis juliflora (PJ), Rhizopus nigricans (RN), and wheat dust (WD), were taken as representatives of various groups of allergens viz, pollen, fungi and dust. The extracts were stored in buffer containing phenol (0.4%) or glycerol (50%) at temperatures ranging from 4-55 degrees C for 15 to 60 days. Protein content of PJ extract was reduced remarkably when it was stored at 40 degrees C for 45 days. Thin layer isoelectric focusing and rocket immunoelectrophoresis of PJ showed that certain antigenic proteins degrade rapidly even at 25 degrees C as early as day 15. However, two to three proteins of PJ remain stable at a higher temperature (40 degrees C) for two months. Relative radioallergosorbent test (RAST) inhibition showed substantial loss of allergenic activity in all the three extracts, when stored at higher temperatures (25-55 degrees C) even for short durations, i.e., 15 days. Extracts (PJ and RN) containing 50% glycerol were found to be stable, retaining more than 50% activity, even when stored at 55 degrees C for 40 days, while extracts without glycerol lost more than 75% of their allergenic activity. However, addition of glycerol did not change the stability of wheat dust allergenic extract. The present findings indicate that allergenic extracts behave differently when stored. Hence, the stability of each extract should be determined individually.

Allergens↗

Allergenic components of a novel food, Micronesian nut Nangai (Canarium indicum), shows IgE cross-reactivity in pollen allergic patients.

BACKGROUND: New foods may present a risk for food hypersensitive patients. Several examples exist of allergic reactions caused by cross-reactive plant-derived foods, and new foods should be scrutinised before introducing them to the market. We have evaluated the clinical and serological relevance of cross-reactivity between Nangai and pollen allergens. METHODS: Cross-reactivity was examined with Maxisorp RAST (radioallergosorbent test), RAST inhibition and Western blot, using sera from patients allergic to grass, birch and mugwort pollen. None of the patients reported having seen or eaten Nangai previously. To determine the biological and clinical relevance of the cross-reactivity, histamine release (HR) test, skin prick test (SPT) and food challenge were used. RESULTS: There was prevalence for reactivity against Nangai in the group of pollen allergic patients. This cross-reactivity seems to be related--at least in part--to carbohydrate epitopes. Three out of 12 patients tested with Nangai were positive upon open challenge, but using double blind placebo controlled food challenge (DBPCFC) this could not be confirmed in two patients. The biological effects of Nangai on allergic patients were confirmed using HR and SPT. CONCLUSION: The Nangai specific IgE found among pollen allergic patients addresses the need for control of new or changed foods before introduction to the market.

Allergens↗

Evaluation and risk factors of latex allergy in spina bifida patients: is it preventable?

Allergic reactions to latex, varying from urticaria to life-threatening IgE-mediated anaphylaxis, have been described in patients with spina bifida. In a study of 50 patients 60% had a latex allergy identified by history, a radioallergosorbent test and/or a skin prick test. Latex allergic patients had undergone statistically more surgical procedures than nonallergic patients (9.5 versus 6.7) with a p value of 0.03. The presence of latex allergy did not correlate with the number of intra-abdominal procedures (1.9 versus 1.4) or with the number of years on clean intermittent catheterization (7.7 versus 5.9). These results suggest that latex allergy is acquired through multiple exposures due to breakdown of blood tissue barriers. Therefore, we recommend that all patients with spina bifida undergoing surgery be evaluated for latex allergy and that all surgical procedures in spina bifida patients, from the time of birth, be performed in a latex-free environment.

Adolescent↗

Demonstration of allergen components in the storage mite Lepidoglyphus destructor by an immunoblotting technique.

The allergen composition of crude extracts of the storage mite Lepidoglyphus destructor were investigated by sodium dodecylsulphate polyacrylamide gel electrophoresis and immunoblotting. The allergen components were detected by sera from allergic farmers with a positive radioallergosorbent test to L. destructor. The allergen-antibody complexes were visualized by rabbit anti-IgE and beta-galactosidase-labelled sheep antirabbit IgG in a chromogenic substrate. Six out of the forty-three sera also had IgE antibodies to the culture medium, which were neutralized by addition of medium antigens before the demonstration of mite allergen components. A total of 11 allergen components were identified. Two of them were found to be major allergen components, identified by more than 50% of the 43 sera used in the study. These 2 IgE-binding components had molecular weights of approximately 18,000 and 16,000 daltons, respectively.

Allergens↗

IgE antibodies to human growth hormone prior to and during treatment.

Clinical manifestations of allergy to biosynthetic hGH were not reported in 245 patients treated for one year. A sensitive radioallergosorbent test showed, however, the presence of anti-hGH IgE antibodies in the sera of 13%. All patients with anti-hGH IgE had also elevated concentrations of total serum IgE, similar to atopic persons (2459 +/- 147 micrograms/l, normal 70 +/- 8, 2p less than 0.0001); 83% of all patients who had specific IgE antibodies to hGH had specific antibodies prior to treatment. Administration of hGH did not consistently change concentrations (up to 8 micrograms/l) or affinities (less than 10(7) to 1.3 X 10(9) l/mol) of the specific reagins. Anti-hGH IgE antibodies were related to atopy. Treatment with biosynthetic hGH did not significantly stimulate specific IgE.

Adolescent↗

The role of IgE-mediated hypersensitivity in otitis media with effusion.

INTRODUCTION: The role of immunoglobin (Ig) E-mediated hypersensitivity in otitis media with effusion (OME) has been the subject of much controversy and personal bias. Uncontrolled studies report the incidence of respiratory allergy in children with OME to range from 4% to over 90%. MATERIALS AND METHODS: Children 1 to 18 years old who had OME requiring a myringotomy and tubes from January 1, 1987, through July 1, 1988, were the subjects of this study. The history of allergic and radioallergosorbent test (RAST) analysis of specific serum IgE levels was undertaken for 26 allergens on 89 children in the study group and 59 children in the control group. RESULT: The incidence of allergen as diagnosed by RAST score was higher in children with OME than children in the control group (P > .05). There was no correlation between positive RAST scores and the number of polyethylene-ventilating (PE) tubes previously placed, history of tonsillectomy, history of adenoidectomy, or the character of the effusion (mucous v serous). CONCLUSION: Respiratory allergy is a factor involved in the pathogenesis of OME.

Adenoidectomy↗

Early diagnosis of infantile seborrhoeic dermatitis and atopic dermatitis--total and specific IgE levels.

Total and antigen-specific IgE levels using the radioallergosorbent test (RAST) were measured in a prospective study of thirty-seven infants with dermatitis of early infancy, with the aim of distinguishing between atopic and infantile seborrhoeic dermatitis at an earlier age than is usually possible on clinical grounds. The results were correlated with the clinical diagnoses made after 1-2 years close follow-up. They showed that a RAST screen to egg white and to milk antibodies, and to a lesser extent total IgE levels, are of value in diagnosing atopic dermatitis at an early age and distinguishing this from infantile seborrhoeic dermatitis.

Dermatitis, Atopic↗