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PubMed1989

Maxisorp RAST. A sensitive method for detection of antigen-specific human IgE in culture fluids.

For determination of allergen-specific IgE in cell culture supernatants and other highly diluted IgE preparations a radioallergosorbent test (RAST) based on high adsorption polystyrene test tubes has been developed ("Maxisorp RAST"). Cladosporium herbarum extract was used as a model allergen but timothy grass pollen, house dust mite and dog dander showed similar results. The test showed specificities of both allergen and immunoglobulin isotype and significant correlations (r = 0.67-0.88) with established RAST procedures were found. Based on immunosorbent-purified allergen-specific IgE the estimated sensitivity was within the order of 150-300 pg allergen-specific IgE per ml. The within-assay variation was 4-9% and the inter-assay-variation 17-29%. The Maxisorp RAST is useful as an inhibition assay for quantitating allergenic activity down to 0.1 biological units/ml of allergen extracts.

Allergens↗

PubMed1990

Diagnostic value of a glass fibre-based histamine analysis for allergy testing in children.

The aim of this study was to compare the diagnostic value of common allergy tests with basophil histamine release in 124 children with symptoms of asthma. The patients were evaluated by case history, skin prick test, RAST-analysis, and basophil histamine release using a glass fibre-based histamine assay to 10 common inhalant allergens. The bronchial provocation test was used as a reference of "true" IgE-mediated asthma. To compare the various diagnostic parameters each absolute test value was classified into a scoring system. The concordance between the tests varied between 85-97%. In general, the best concordance was found between basophil histamine release and RAST. Sensitivity, specificity and predictive values were calculated on the basis of 104 bronchial provocation tests. It was found that histamine release was the best single analysis, followed by RAST and prick testing. The sensitivity of RAST and histamine release was very high (1.00) for pollen and house dust mites. Histamine release showed a predictive value between 0.91 and 1.00 for pollen and house dust mites, thus indicating the possibility of omitting the bronchial provocation test. In the dander group histamine release gave the best sensitivity (0.91), however at the expense of specificity (0.64), whereas RAST and skin prick test gave a specificity of 1.00. In the mould group histamine release also showed the best diagnostic value. The combination of skin testing with histamine release or RAST was of no additional diagnostic help. It is concluded that the glass fibre-based histamine analysis, which makes routine histamine release testing possible, is a reliable diagnostic test in children.

Asthma↗

PubMed1990

Endotoxin from Haemophilus influenzae enhances IgE-mediated and non-immunological histamine release.

Haemophilus influenza and its extracellular products (EP) did not release histamine from basophil leukocytes in cell suspensions from normal individuals, patients with chronic bronchitis or patients allergic to either house dust mite, grass pollen, cat dander or to their own bacteria. However, the EP was found to enhance their basophil histamine release. IgE-mediated histamine release was examined by stimulation of the cells with anti-IgE or the specific allergens, and non-immunological histamine release by stimulating the cells with the calcium ionophore A23187 or Staphylococcus aureus. In all the experiments EP caused a significant increase in the histamine release. When H. influenzae endotoxins were removed from the EP, the potentiating effect of EP was completely abolished, whereas heating (80 degrees C, 30 min) or treatment of EP with proteinase did not influence the potentiating effect. These results indicate that H. influenzae endotoxin potentiates histamine release caused by IgE-mediated reactions or by non-immunological mechanisms.

Adult↗

PubMed1990

Passive sensitization and histamine release of basophils. IgE and cellular factors regulating histamine release.

This study had two purposes. First, to examine a possible functional heterogeneity of IgE regulating basophil histamine release and the effect of using two different donor cells for passive sensitization experiments. Second, to investigate basophils not releasing histamine to anti-IgE by stimulating protein kinase C with the addition of the phorbol-ester, TPA. In consecutive experiments responding donor basophils were passively sensitized with plasma from non-responding subjects. Thus, the first set of experiments included passive sensitization of acid treated donor basophils from one atopic and one non-atopic patient with plasma from 29 children with exogenous asthma to grass pollen, cat dander, or dust mites. Different secretagogues (anti-IgE, Concanavalin A, and N-formyl-methionyl-leucyl-phenylalanine) induced different histamine release responses due to a cellular property of the basophils not related to the type of IgE bound to the cell membrane. It was demonstrated that the allergen-induced histamine release did not depend on the extract or type of IgE when the biological activity of each extract and serum-specific IgE levels were similar. However, the atopic donor cells released significantly (P less than 0.05) more histamine than non-atopic donor cells. Thus, histamine release depends on the type of secretagogues and a cellular property which is maybe influenced by the presence of serum factors and a certain type of IgE in the serum of atopics. The second set of experiments included 10 patients (6 atopics and 4 non-atopics) with non-histamine releasing basophils. In the presence of 10 ng/ml TPA, however, seven of 10 patients released histamine at anti-IgE challenge. Three months later two additional patients became responsive in the presence of TPA. By passive sensitization of responding donor basophils the non-responding patients were shown to possess functionally intact IgE. Thus, the discrepancies sometimes observed between clinical symptoms, serological IgE-antibody measurements and histamine release testing in allergic patients may be related to a cellular property of basophils.

Adolescent↗

PubMed1991

Severe anaphylactic reaction to thymostimulin.

Thymostimulin is a partially purified extract of calf thymus, consisting of a group of polypeptides with a total molecular weight of approximately 12,000 D. It is used as an immunologic response modifier in primary and secondary immunodeficiencies. We report a patient who had a severe anaphylactic reaction with the first injection of thymostimulin. Type I hypersensitivity to thymostimulin was suggested by an immediate skin test response, specific IgE determination by indirect ELISA and a positive histamine release test. This patient also reacted to bovine serum albumin (BSA), fetal calf serum (FCS) and cow hair and dander. The controls did not react to any of these tests. Clinicians need to be aware that the use of medical products of xenogeneic origin may lead to severe allergic reactions.

Adjuvants, Immunologic↗

PubMed1992

A case of fallow deer allergy. Cross-reactivity between fallow deer and horse allergy.

We report the case of a 25-year-old housewife, a native of Mexico City, with rhinitis and asthma induced by exposure to fallow deer in an animal park close to her home. In the literature, we could find only one previous case. The patient presented a polyvalent IgE sensitization in prick skin tests and RAST to several animals' dander and epithelia, but RAST inhibition experiments showed a cross-reactivity only between fallow deer and horse allergen extracts.

Adult↗

PubMed1992

Natural history of bronchial asthma in childhood. A prospective study from birth up to 12-14 years of age.

In a cohort of 1654 consecutively born children followed from birth, the cumulated incidence of asthma up to 11 years was 5.3% and the prevalence 3% between 10 and 11 years of age. The asthma début was prior to 1.5 years in 1/3 and before 3 years in 1/2 of the children. Among 59 children examined at 11.5-14.5 years of age, 16 (27%) had no prevalent asthma. On the other hand, inadequate medication was found in 13, undiagnosed chest deformity in four, and wheezing in seven children. All 54 tested children including those with no prevalent asthma had a PC20 histamine less than 8 mg/ml. Tread mill test gave a significant reaction in 15 children. Animal danders and pollens were the most common allergens giving reactions at skin prick test. Compared to one earlier Swedish study an increase in mite sensitivity was found. Cord-blood IgE concentration and a positive immediate family history of atopic disease had no predictive value for the severity of asthma at this follow-up. A new total asthma score including number of days with functional impact of asthma during the last year and present medication was used for group comparisons. Children with a high score at 11.5-14.5 years had more exercise-induced asthma, more concomitant allergic diseases, earlier asthma start, more chest deformity, hyperreactivity both on tread mill and histamine challenge tests, elevated IgE, positive Phadiatop and more reactions at skin prick test, especially to animals and mites. Bronchial asthma was found more often in children born during August through October, possibly due to unsuitable indoor climate and more virus infections during their first 6 months of life. The severity of the asthma was, however, not influenced by the month of birth. No significant differences were found between boys and girls regarding the age at début, asthma severity at follow-up, or bronchial histamine threshold levels.

Adolescent↗

PubMed1992

Age-dependency of sensitization to aero-allergens in asthmatics.

Skin reactivity (intracutaneous test) to histamine and allergens was studied cross-sectionally in a Dutch asthmatic patient population from childhood to old age (4-75 years). It was found that the histamine skin reactivity rose significantly (p less than 0.05) during childhood, was significantly higher in the 10-15-year age group, and was constant between 20 and 75 years of age. The mean wheal index (histamine ratio) of all allergens was constant during childhood, and decreased after the age of 25 for grass pollen and house-dust mite and after the age of 15 for the other allergens. The prevalence of a positive skin test decreased with age, except for grass pollen. During childhood the indoor allergens, cat dander and house-dust mite, were the most important, while after the age of 15 sensitivity to an outdoor allergen, grass pollen, increased markedly. At all ages house-dust mite was the most important allergen. After the age of 25 the prevalence of every allergen declines. The prevalence of a positive skin test to Cladosporium was unexpectedly high in childhood (10-40%). It can be concluded that the prevalence of a positive skin test declines with age, except for grass pollen. The degree of sensitization in asthmatics peaked in the age groups between 20 and 40 and sensitivity to indoor allergens developed earlier than sensitivity to outdoor allergens.

Adolescent↗

PubMed1993

Skin prick test reactions to brewer's yeast (Saccharomyces cerevisiae) in adult atopic dermatitis patients.

The sensitizing capacity of brewer's yeast (Saccharomyces cerevisiae) was studied with the skin prick test method in 449 subjects, including 226 atopic dermatitis (AD) patients, 50 patients with allergic rhinitis (AR) and/or asthma (A), and 173 nonatopic controls. A positive SPT reaction (> or = + +) was seen in 94% of patients with severe AD, in 76% with moderate AD, and in 25% with mild AD or no history of AD. Patients with AR and/or A and nonatopic controls displayed a positive reaction in only 8 and 2% of cases, respectively. There was also a parallel skin prick test reactivity with other yeasts including Pityrosporum ovale and Candida albicans, suggesting cross-reactivity. Parallel skin reactivity was observed also with molds and animal dander but not with pollen or house-dust mite. A significant correlation was also found between total serum IgE level and skin prick test (SPT) results with S. cerevisiae.

Adolescent↗

PubMed1993

Optimization of skin testing. II. Evaluation of concentration and cutoff values, as compared with RAST and clinical history, in a multicenter study.

In this multicenter study we evaluated the results of a previous study (part I) in a relatively large Dutch patient population, using the two previously tested allergens (house-dust mite and grass pollen) and two other standardized allergens (tree pollen and cat dander). The obtained skin test results were expressed as a histamine ratio and compared with RAST and clinical history (CH). The sensitivity and specificity were calculated at different cutoff values of the skin tests. The optimum cutoff values of 0.7 intracutaneous tests (ICT) and 0.4 skin prick tests (SPT) resulted in a predictive value for the detection of allergic sensitization of 83% (RAST) and 77% (CH), and 91% (RAST) and 86% (CH), for the ICT and SPT, respectively. As the ICT and SPT were performed in different centers, the results of these methods cannot be compared. No systemic side-effects of the skin tests were recorded. These results generally correspond well with the predictions regarding safety and predictive value of part I of this study, in which a limited number of patients was studied. In conclusion, through the use of a limited number of standardized allergens in a small group of patients, it may be possible to predict a safe and efficacious concentration for routine skin testing and to extrapolate from these results to other standardized allergens.

Adolescent↗

PubMed1993

In vitro diagnosis of farmers' IgE-mediated allergy by Phadiatop and three new multiallergen RAST analyses.

Identification of persons with atopic allergy in a population is difficult if the pattern of sensitizing allergens is unique. Three new multiallergen tests, Professional Allergy Mixes (PAX) 1-3 (Pharmacia Diagnostics, Uppsala, Sweden), have recently been introduced to screen IgE-mediated allergy among farmers. PAX 1 contains animal danders and feathers, PAX 2 mites and insects, and PAX 3 pollens and molds. The value of these new tests for the determination of IgE-mediated allergy among farmers was compared with that of Phadiatop, a test for general atopy. Of 440 farmers, 91 were positive to Phadiatop and 97 to any of PAX 1-3. None of the farmers were positive in all tests. Only 70/97 sera positive in RAST to PAX 1-3 had a positive Phadiatop, whereas of 91 sera positive in Phadiatop, 70 had specific IgE antibodies to PAX 1-3. The value of using either Phadiatop or PAX 1-3 is limited (sensitivity 74-79%), but, together, these four tests will detect almost all (95.9%) farmers with IgE-mediated sensitization and all (100%) with symptoms. We have also shown that the grain weevil Sitophilus granarius and the botfly Gasterophilus intestinalis are important sources of allergens in rural environments.

Agricultural Workers' Diseases↗

PubMed1993

The batch-to-batch variation of the potency of dog- and cat-allergen-coated lancets. Evaluation by skin prick testing.

In clinical work I formed the impression that the potency of Phazet lancets, for cat and dog allergens, was lower than earlier and too low to diagnose all clinically sensitive children with cat and dog allergy. Therefore, I decided to investigate the potency of newly produced and several years old Phazet lancets coated with dog and cat allergens. Twenty-six adults with case histories of cat and/or dog allergy and skin reactive to extracts of either dog dander or cat epithelium were skin prick tested in duplicate with one old batch and one newly produced batch of Phazet dog and/or cat lancets as well as the in-house reference of cat and/or dog allergen, 100,000 BU/ml. The potency of Phazet lancets in relation to the standard was evaluated by the median slope of the allergen dose-response relationship. The potency of the old dog lancets (mean 48,900 BU/ml) was higher (NS) than that of the newly produced lancets (mean 28,000 BU/ml), and the potency of both types of lancets was significantly less (P < 0.001) than that of the dog allergen in-house standard. On the other hand, the potency of newly produced (123,000 BU/ml), but not old (108,000), cat-allergen-coated Phazet lancets was higher than that of the cat allergen in-house reference extract (P = 0.048). There was a marked variation in potency between patients. Most of this variation was due to factors varying between patients, not between lancets, indicating that the composition of the allergen on the lancets was partly different from that of the standard.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1994

Optimization and characterization of freeze-dried multilamellar liposomes incorporating different standardized allergen extracts.

Desensitization therapy for type I allergy is now current practice. Liposomes have been proposed as a support for allergens to improve safety and effectiveness. The aim of this work was to optimize liposomal formulations of three different standardized allergen extracts and to test their allergenicity in vitro and in a preclinical trial. Allergen extracts (Dactylis glomerata, Dermatophagoides pteronyssinus, and cat hair and dander) were associated with multilamellar liposomes of varying compositions at different pH. Liposome-bound allergens were quantified by RAST inhibition after ultracentrifugation, and analyzed qualitatively by SDS-PAGE followed by immunoblotting. Their allergenicity was assessed by basophil degranulation in vitro, as compared with the allergenicity of an aqueous extract, and by skin tests in allergic subjects. The best association, about 50% of the added allergen, was obtained with negatively charged liposomes, when the pH of the allergen solution was adjusted so as to impart a net positive charge to the proteins. One-third of the liposome-associated allergens was located on the surface of the liposomes and was free to interact with antibody, as shown by RAST inhibition assays and the basophil degranulation test; the remaining two-thirds was encapsulated within the liposomes. All the major immunoreactive proteins in the extract were included. These liposomes could be readily freeze-dried and reconstituted without changing their properties. This study reveals the allergenic characteristics of liposomes and suggests their potential use in the treatment of allergic patients.

Allergens↗

PubMed1994

Prediction of sensitization to flour allergens.

The purpose of the present study was to investigate the factors that may contribute to the sensitization to flour in baker's apprentices, and to predict this type of sensitization with a stepwise logistic regression model. Of the total of 258 baker's apprentices, 54 (20.9%) exhibited specific IgE development against flour allergens (wheat, rye, and/or barley). Allergologic variables, such as the total IgE level and IgE specific to grass pollen, tree pollen, weed pollen, molds, and animal dander, were found to correlate strongly with the sensitization to flour. However, the correlation between the presence of other manifest allergies and the sensitization to flour was weak. The presence of shadowing of the paranasal sinuses on the radiograph (mucosal thickening, opacity, air/fluid levels, and/or polypous shadowing) correlated with flour sensitization as well as with the leukocyte and lymphocyte counts in peripheral blood. Of the lung function data, only total lung capacity (in percentage of the predicted values) weakly correlated with the sensitization to flour. We were able to predict flour sensitization with a sensitivity of 63%, a specificity of 90%, and an efficiency of 84% by using the parameters that allow for the presence or absence of IgE specific to grass pollen and tree pollen, the presence or absence of radiographic shadowing, and total lung capacity in percentage of the predicted values.

Allergens↗

PubMed1995

Causative allergens of allergic rhinitis in Japan with special reference to silkworm moth allergen.

To determine the importance of silkworm moth allergens, we tested 267 patients with allergic rhinitis with CAP-RAST, a new assay system for detecting specific IgE in sera. We used the following allergen sources: house-dust mite, Candida, Alternaria, cat dander, orchard grass, ragweed, mugwort, Japanese hops, Japanese cedar, Japanese cypress, cotton, silk, larval chironomid midge, adult chironomid midge, and silkworm moth. As expected, the highest positive rate of reaction, as determined by CAP-RAST, was 73.8% for Japanese cedar, followed by 53.2% for mite and 50.6% for Japanese cypress. Although the positive rate for silkworm moth was not as high as for cedar pollen, one-third of patients had specific IgE against silkworm moth. The positive rate of reaction to silkworm moth was much higher than that to chironomid. It was interesting to note that the patients reported had not been documented as having frequent contact with silkworm moth allergen. Correlations between silkworm moth and silk, and silkworm moth and chironomid midge were found to be significant (P < 0.001). Silkworm moth allergen showed the third highest reaction rate in patients with severe symptoms. These results suggest that silkworm moth allergen should be considered to be important in patients with allergic rhinitis in Japan.

Adolescent↗

PubMed1995

Severe anaphylactic reaction to bovine serum albumin at the first attempt of artificial insemination.

A 33-year-old woman without history of previous atopic diseases or drug allergies developed a severe anaphylactic reaction with asthma, vomiting, itching, generalized urticaria, and angioedema during artificial insemination with her husband's sperm. The sperm-processing medium contained bovine serum albumin (BSA). Skin prick test and RAST demonstrated an IgE-mediated hypersensitivity to BSA as well as a polyvalent atopic sensitization to pollens, animal danders, cow's milk, beef, pork, and mutton. SDS-PAGE studies indicated serum albumin to be the appropriate allergen with a high degree of cross-reactivity between serum albumin from different animal species. Artificial insemination with fluid containing potential allergens can, therefore, represent an unnecessary risk for atopic females, even in the absence of prior clinical symptoms of allergic diseases. Preoperative testing with the medium is recommended.

Adult↗

PubMed1997

Occupational type I allergy to Christmas cactus (Schlumbergera).

The study aimed to determine whether occupational contact urticaria and symptoms of mucous membranes, reported by five workers in a cactus nursery, were due to IgE-mediated allergy to Schlumbergera cacti. The five persons had positive skin prick tests to the plants as is and positive histamine-release tests, and in three of them specific IgE to the cacti could be demonstrated by Maxisorp RAST and immunoblotting. Four of the patients were atopic, and the fifth had a positive skin prick test to cat dander, indicating latent atopy. Skin prick tests with cacti were negative in most atopic volunteers, and all had negative histamine-release tests. The results suggest a true IgE-mediated allergy to the cacti, and both genetic predisposition and close contact with the plants at work seem to be important factors in the emergence of this new occupational allergy.

Adult↗

PubMed1998

Deposition of cat (Fel d 1), dog (Can f 1), and horse allergen over time in public environments--a model of dispersion.

BACKGROUND: The occurrence and accumulation over time in public environments of cat, dog, and horse allergens was evaluated. METHODS: Concentrations of animal danders were analyzed by ELISA and countercurrent immunoelectrophoresis (CCIE). RESULTS: Among factory-new mattresses, 15/17 contained detectable levels of cat and/or dog allergen, whereas no horse allergen was found although six of the mattresses were stuffed with horsehair. Dust from 15 used mattresses contained significantly higher concentrations of Fe1 d 1 and Can f 1 than the factory-new ones (P < 0.001). Allergen concentrations and titers correlated to the period of time that the mattresses had been tried by customers; rs = 0.52-0.77, P = 0.04-0.001 (cat), rs = 0.38-0.48, P = 0.15-0.08 (dog), and rs = 0.64-0.74, P = 0.008-0.003 (horse). The increase over time occurred rapidly in highly frequented stores and after 3 weeks reached concentrations that have been found in homes where furred pets had formerly been kept or even the lower allergen scale of homes where pets were currently kept. CONCLUSIONS: The dispersion of allergens from furred animals to pet-free public places is likely to occur by deposition from people who have been in direct or indirect contact with pets, and high levels of such allergens seem to accumulate in a short period of time.

Allergens↗