Quantification of the major allergen from cypress (Cupressus arizonica) pollen, Cup a 1, by monoclonal antibody-based ELISA.

BACKGROUND: Cypress pollen allergy is an important cause of rhinoconjunctivitis and asthma in Mediterranean countries. Cypress allergenic extracts are difficult to produce since they have low protein and high carbohydrate content, thus accurate standardization of them is essential to guarantee their quality. The aim of this study is to develop a sandwich ELISA for the quantification of Cup a 1, the major allergen of cypress (Cupressus arizonica) pollen extract. METHODS: Monoclonal antibodies directed to purified Cup a 1 were produced. Two of them (9C7 as capture antibody and 3D2 as the tracer) were selected to develop a quantitative sandwich ELISA. This ELISA was subsequently evaluated and compared with other techniques. RESULTS: The described ELISA is very sensitive with a detection limit of 8.7 ng/ml and a practical working range of 62.5-1,000 ng/ml. The assay is also highly reproducible with intra-assay and interassay coefficients of variation of less than 10%. The purified Cup a 1, used as standard, presents pectate lyase enzymatic activity. The assay also detected Cup a 1-like proteins in pollen from other Cupressaceae. A good correlation was obtained between Cup a 1 content of 12 C. arizonica pollen extracts and their IgE-binding activity. CONCLUSIONS: The described Cup a 1 ELISA is sensitive, specific and reproducible and can be used for the quantification of Cup a 1 in C. arizonica and other related pollen extracts. It also provides a reliable indication of the allergenic activity of the whole cypress pollen extract.

Hypersensitivity pneumonitis in workers exposed to esparto grass (Stipa tenacissima) fibers.

Esparto grass (Stipa tenacissima), which is commonly found in the Mediterranean countries, has a wide variety of uses. Five stucco makers who had cough, dyspnea, malaise, and fever after exposure to esparto fiber used in their jobs showed a significant decrease in symptoms when they were away from work. Precipitating antibodies against an esparto extract were found in the sera of all patients. Specific IgG antibodies against the esparto extract were also demonstrated in all patient sera, as were IgG antibodies to Aspergillus fumigatus and thermophilic microorganisms (Micropolyspora faeni and Thermoactinomyces vulgaris) by means of an ELISA method. Esparto activity was inhibited in different ranges by the above antigens by inhibition ELISA. Only A. fumigatus could be identified after microbiologic evaluation of the esparto fiber samples. After inhalation challenge tests were performed with esparto extracts, all patients showed significant decreases in forced vital capacity, transfer lung CO, and PaO2 blood gas from baseline values. Fever, chills, malaise, dry cough, tachycardia, tachypnea, and rales on chest auscultation were also observed in all patients. Findings from bronchoalveolar lavage were suggestive of allergic alveolitis. Transbronchial biopsy specimens showed interstitial alveolitis with lymphocyte-macrophage infiltrate and granuloma. Unexposed control subjects did not exhibit reactivity to any of the tests listed above. The dust derived from esparto fibers can cause hypersensitivity pneumonitis in exposed subjects. Organisms such as A. fumigatus and thermophilic actinomyces could be the causative antigens. "Stipatosis" might be an appropriate name for this disorder.

[Multicenter drug surveillance of sublingual immunotherapy in allergic patients]. / Estudio multicéntrico de farmacovigilancia de inmunoterapia sublingual en pacientes alérgicos.

A study of pharmacovigilance has been performed in 522 patients suffering from allergic rhinitis and/or asthma. Sensitization was due to pollens and mites in ther majority of cases. The treatment consisted in biologically standardized glycerinated allergenic extracts for specific sublingual immunotherapy (SLIT) and, major allergens were also quantified, depending on the allergenic composition. The aims of the study were to assay the effect of the SLIT in a short-and-medium-term course and to know the adverse reactions most frequently appearing, as well as the unexpected side effects that are only possible to know through the study of a numerous sample of patients. We also evaluated the factors that can modify the tolerance of the treatment, such as the onset of patient's disease and its severity. All the patients had a clinical history of, at least, two years of evolution and positive skin-prick test to the allergen/s cause of their atopic disorder. Exclusion criteria were immunotherapy in the last two years, pregnancy and those situations in which the immunotherapy is contraindicated according to the EAACI requirements. In order to carry out the daily record of the SLIT as well as the possible appearance of symptoms, two diaries were given to each patient. Tolerance was evaluated by the physician at the end of the build-up phase (3 months) and when the maintenance phase finished (5 months later). Total administered doses were 44.021, the final number of patients who registered any adverse reaction in relation with SLIT was 41 (7.9%), with a number of 67 adverse reactions that means a 0.15% over the total administered doses.(ABSTRACT TRUNCATED AT 250 WORDS)

Green bean hypersensitivity: an occupational allergy in a homemaker.

As a member of the legume family, the green bean is frequently associated with food allergy. However, allergic reactions caused by skin contact or by inhalation of vapors from boiling legumes are rare. This article presents a case of occupational asthma in a homemaker; symptoms occurred during preparation and cooking of raw green beans. Skin prick, rub, and bronchial provocation tests were performed on the patient. In vitro tests were done with the serum samples of the patient and 10 control subjects (5 atopic and 5 nonatopic). Test results indicate that the patient has type I hypersensitivity to raw green bean antigen(s). This case is of interest because it demonstrates that a food allergen, when inhaled, can induce respiratory symptoms in sensitized patients and may even be the source of primary sensitization.

Bronchial asthma induced by chick pea and lentil.

Allergic reactions to legumes through inhalation have rarely been described. We report the case of a 20-year-old man who experienced asthmatic attacks when exposed to the steam from cooking either chick pea or lentil. Type I hypersensitivity to the antigens in these legumes was demonstrated by means of immediate skin reactivity, histamine release tests, RAST and RAST inhibition. Specific bronchial challenges with the heated (75 degrees for 30 min) extracts of chick pea and lentil elicited isolated immediate responses.

Swiss chard hypersensitivity: clinical and immunologic study.

Allergy to vegetables and fruits seems to be more prevalent in atopics, especially in birch pollen-sensitized individuals. We report a case of a grass pollen-sensitized woman, in whom the inhalation of vapor from boiling Swiss chard precipitated rhinoconjunctivitis and asthma. Type I hypersensitivity to Swiss chard was demonstrated by means of immediate skin test reactivity, specific IgE determination by RAST, basophil degranulation, histamine release test, and an immediate bronchial provocation test response to Swiss chard extract. The controls did not react to any of these tests. RAST inhibition assays suggest the presence of some cross-reactivity among Swiss chard and grass pollen antigens, as well as cross-reactivity between vegetables and weed pollens of the chenopod family.

Effect of nifedipine on skin prick tests.

The immediate skin test due to interaction between allergen and mast cell bound IgE is one of the cornerstones in the clinical allergy workup. The release of histamine and other mediators from basophils and mast cells depends on the influx of Ca2+ into these cells when stimulated. The aim of this study was to evaluate the effect of common therapeutic doses of nifedipine (NFD), one of the calcium channel blockers, on the allergen skin tests. We prick tested 23 grass sensitive individuals with 7 different grass pollens at three times: at basal conditions (T0), 30 min. after having taken 20 mg of NFD s. l. (T1), and 17 of them after a week of receiving twice a day 20 mg of a NFD retard form (T2). The wheal surface obtained for each substance (allergen, histamine) at T0 was considered as basal value and compared with the one obtained at T1 and T2 for the same substance by the Wilcoxon's test. We found a significant increase in the wheal surfaces, both with allergen and histamine, at T1 and T2. In contrast to what could be expected, common therapeutic doses of NFD produce a discrete but statistically significant increase of the PT. Factors such as arteriolar vasodilation could be implicated. The increase of the allergen prick test and the increase of the histamine prick test both at T1 and T2 were not statistically different. Therefore, we do not think it necessary to stop NFD before allergen skin testing.