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PubMed1994

Measurement of allergen-specific IgD and correlation with allergen-specific IgE.

A new method for the measurement of allergen-specific IgD (as-IgD) was developed by modifying the ImmunoCAP assay (Pharmacia), and amplification of the signal with a goat anti-human/rabbit anti-goat detection system. The assay was sensitive enough to measure as-IgD in serum samples. The specificity of the assay was examined using inhibition tests with excess corresponding and non-corresponding allergens. For the different allergens inhibition rates between 56% (house dust mite) and 88% (cat) could be achieved. Non-corresponding allergens did not inhibit the as-IgD binding. Total IgE and allergen-specific IgE (as-IgE) was measured using the ImmunoCAP system. Total IgD was measured using a sandwich ELISA. As-IgD was measured in serum samples from 51 atopic and 23 non-atopic subjects, and the correlation with as-IgE was examined. As-IgD was detected in both atopics and non-atopics but at higher levels in atopics. As-IgD against birch pollen and timothy pollen allergen was found to be increased in atopics with IgE directed against these allergens compared to atopics without IgE against these allergens (P < 0.02 and P < 0.03). As-IgD against birch pollen allergen was higher in atopics with IgE specific to this allergen than in non-atopics (P < 0.02). In contrast to total IgE and total IgD, significant correlations were observed between as-IgD and as-IgE against timothy pollen (r = 0.34, P < 0.04), birch pollen (r = 0.38, P < 0.05) and cat dander allergen (r = 0.52, P < 0.01). The observed correlations between as-IgD and IgE suggest that IgD and IgE may be similarly regulated, and thus the measurement of as-IgD may give further insight into the regulation of IgE.

Adolescent↗

PubMed2006

Evaluation of three different histamine concentrations in intradermal testing of normal cats and attempted determination of 'irritant' threshold concentrations for 48 allergens.

The purpose of this study was to determine the optimal histamine concentration and 'irritant' allergen threshold concentrations in intradermal testing (IDT) in normal cats. Thirty healthy cats were tested with three different histamine concentrations and four different concentrations of each allergen. The optimal histamine concentration was determined to be 1: 50,000 w/v (0.05 mg mL(-1)). Using this histamine concentration, the 'irritant' threshold concentration for most allergens was above the highest concentrations tested (4,000 PNU mL(-1) for 41 allergens and 700 PNU mL(-1) for human dander). The 'irritant' threshold concentration for flea antigen was determined to be 1:750 w/v. More than 10% of the tested cats showed positive reactions to Dermatophagoides farinae, Dermatophagoides pteronyssinus, housefly, mosquito and moth at every allergen concentration, which suggests that the 'irritant' threshold concentration for these allergens is below 1,000 PNU mL(-1), the lowest allergen concentration tested. Our results confirm previous studies in indicating that allergen and histamine concentrations used in feline IDT may need to be revised.

Allergens↗

PubMed1979

3H-histamine release from human leukocytes. A new method for detecting type I allergy compared with basophil histamine release technique.

A rapid, simple, and inexpensive method for large scale screening of patients suspected of type I allergy has been developed. The method is based on in vitro incorporation of 3H-histamine in the leukocytes of the patient, whereafter release of labelled histamine is measured after provocation of the cells with the suspected allergen. The new method was compared with the conventional basophil histamine release technique by in vitro provocation of six asthmatic patients under suspicion of type I allergy against animal dander, house dust, and mite, and an almost identical release of histamine was observed in both assays.

Allergens↗

PubMed1979

Detection and quantitation of Dermatophagoides antigens in house dust by immunochemical techniques.

Rabbit antibodies against D. pteronyssinus whole culture extract were rendered specific to mite antigens by immunoabsorption with extract of human dander. Using the absorbed antibodies, an extract of a mite culture gave rise to 15 peaks in crossed immunoelectrophoresis and to a titer of 512 in counter-immunoelectrophoresis. When applied to dust samples from the homes of allergic patients, counterimmunoelectrophoresis gave reproducible results within an error of 1 titer-step. Mite antigens were detected in 64% of 105 dust samples investigated. The titer correlated well (P less than 0.005) with the concentration of Dermatophagoides bodies, found in 13 mattress dust samples by microscopy. Advantages and limitations of immunochemical quantitations of house dust mites in dust samples is discussed in relation to investigations by microscopy.

Antibodies↗

PubMed1980

Influence of the pollen season on immediate skin test reactivity to common allergens.

We studied the reproducibility of skin prick test reactions and tested 65 young men, picked from a population survey, in spring and fall. We used 16 extracts from two manufacturers with 12 common allergens. The extracts employed on both occasions were from the same allergen batches and the testing procedure was the same. Four subjects who reacted negatively in spring turned positive when re-tested in fall; the opposite happened to one subject. The result remained positive or negative on both occasions in 92%. The best reproducibility (96%) was obtained by pollens. The reactivity against tree and grass pollens was higher in fall than in spring; this is obviously a result of the immunologic stimuli induced by exposure to pollens during the season. The reactions caused by animal danders and house dust were, however, smaller in fall than in spring. This is believed to be due to young people being less exposed to indoor dust in summer. The immediate skin test reactivity is influenced by several factors; we would like to add one: the time of year when the test is applied. The reproducibility of skin prick test results is quite satifactory when the problem is approached as presented here.

Adolescent↗

PubMed1980

Effect in man of oral terbutaline on cutaneous reactions induced by allergen and cold stimulation.

In eight atopic subjects wheal and flare responses to intradermally injected horse dander and histamine were determined after pretreatment with 5 mg oral terbutaline or placebo in a double-blind cross-over study. In each individual a dose of allergen was used that produced a flare reaction approximately the size of the ED50 for histamine. Pretreatment with terbutaline was found to attenuate both the wheal and the flare reactions to allergen throughout the observation period of 150 min but only the effect on the wheal response reached statistical significance (P less than 0.01). The responses to histamine were not influenced. In five subjects with cold urticaria, treatment with 2.5 mg terbutaline t.i.d. for a week had no effect on the time period of cold provocation needed to evoke an urticarial lesion. It is concluded that oral treatment with terbutaline may produce an inhibitory action on allergen induced reactions but that this effect is not strong enough to interfere with clinical skin testing and hence the drug need not to be withdrawn prior to such testing.

Administration, Oral↗

PubMed1980

Cutaneous responses to allergen after local pretreatment with beta-adrenoceptor stimulating and blocking agents.

beta-Adrenoceptor stimulating agents possess anti-allergic effects in vitro and in vivo. To study the mechanism of action further we compared in 10 atopic subjects the effects on allergen-induced cutaneous reactions of intradermally (i.d.) injected prenalterol (1-10 microgram) terbutaline (2.5-250 ng) and KWD 2131 (100 ng-2.5 microgram), i.e. compounds with preferential actions on beta 1-, beta 2-adrenoceptors and on beta-adrenoceptors not fitting this classification. All injections were given according to a double-blind design. Terbutaline and KWD 2131 produced a dose-dependent inhibition of the skin reactions induced by injecting horse dander allergen 5 min later. Terbutaline was about 20 times as potent as KWD 2131 whereas prenalterol was inactive. Propranolol fully blocked the anti-allergic effect produced by terbutaline. The results suggest that the cutaneous anti-allergic effect of beta-adrenoceptor-stimulating drugs is mediated via activation of beta 2-adrenoceptors, possibly on the mast cell, resulting in inhibition of mediator release.

Adrenergic beta-Agonists↗

PubMed1982

Allergoprints in horse allergic children.

In order to determine the allergenic activity of five purified horse allergens, 22 children allergic to horses according to history, skin test, and leukocyte histamine release were evaluated. Washed leukocytes from all patients were tested for allergen-induced histamine release utilizing four epidermal horse allergens (Ags 6, 9, 11, and 15) and horse serum albumin. Crossed radioimmunoelectrophoresis was carried out with a standard horse dander extract and serum from each patient. The results showed considerable variation in the individual allergoprints. Ag 11 had the highest mean allergenic activity. Sensitivity to horse serum albumin was demonstrated three times. Our data show that the amount of serum IgE antibodies bound by horse allergens correlates significantly with the capacity of the allergens to induce histamine release from washed leukocytes.

Adolescent↗

PubMed1982

Mites living in hay: an important allergen source?

Fifty-two farmers and 21 non-farmers with symptoms related to hay contact were investigated. Skin prick tests (SPT) and RAST were performed with an extract made of barn dust (BDE) consisting of hay remnants. At microscopy, the barn dust was found to contain large amounts of dead mites, most of them identified as Lepidoglyphus destructor and Acarus siro. Twenty-one patients (9 farmers and 12 non-farmers) had positive RAST or BDE, with a good correlation to the case histories. Other allergies, especially to Dermatophagoides pteronyssinus, pollens, and animal danders were also common among patients. However, there was no correlation between positive SPT and RAST to BDE and any other allergen tested. This indicates that BDE contains distinct allergens, most likely of mite origin.

Allergens↗

PubMed1984

A reference allergen preparation of the house dust mite D. pteronyssinus, produced from whole mite culture--a part of the DAS 76 study. Comparison with allergen preparations from other raw materials.

A D. pteronyssinus whole culture allergen preparation contained 49 antigens as revealed by crossed immunoelectrophoresis (CIE), using polyspecific rabbit antibodies. Crossed radio-immunoelectrophoresis ( CRIE ) with sera from 30 patients revealed nine allergens, antigens 42, X, Y and 23 (in rank order) showing the most frequent and intense IgE-uptake. Nine antigens originated from the culture medium (human dander + yeast), but none of these gave rise to specific IgE-uptake. Extremely few and weak reactions were observed in radioallergosorbent (RAST) with 129 sera, using media extracts on the discs. Purified mite body extract (PMB) contained less ag 42 and more ag Y and ag 23 than whole mite culture extract ( WMC ), whereas an acetone-extracted mite excreta preparation (AML) contained 5 times more ag 42, but was devoid of ag Y and ag 23. Ag X was present in all preparations. The RAST-inhibitory potency of PMB was best correlated with the content of ag X. Preparations with properties similar to WMC and PMB were judged as suitable for clinical application.

Allergens↗

PubMed1984

A modified crossed radioimmunoelectrophoretic method for determination of allergen-specific IgG antibodies. Elimination of non-specific binding by use of F(ab')2-fragments of rabbit antibodies and 125I-labelled protein A.

A crossed radioimmunoelectrophoretic (CRIE) method for detection of specific IgG antibodies in patients' sera against horse hair and dander was developed. The unacceptably high non-specific binding encountered when substituting 125I-labelled antihuman IgG for 125I-labelled antihuman IgE in an ordinary CRIE was eliminated by the combined use of 125I-labelled Protein A as detector, and F(ab')2-fragments of the allergen-specific rabbit antibodies. The low background binding thus obtained makes the method useful for detection of specific IgG in sera where the ratio between specific and non-specific IgG is low. Therefore the method should also be applicable to other antigen/allergen systems.

Animals↗

PubMed1986

Histopathological support of antagonism of allergen-induced mast cell mediator release in human skin by a beta-2-agonist.

The site of antagonistic action on allergen-induced early skin reactions by the beta 2-agonist terbutaline was studied by light microscopy in 10 atopic subjects. Pretreatment with 1 microgram terbutaline intradermally 5 min prior to challenge with horse dander allergen produced an approximate inhibition of 85 and 55% of flare and wheal responses respectively (P less than 0.001). Biopsy specimens obtained from skin sites injected with allergen alone showed a reduced number of remaining stainable mast cells as compared to sites injected with terbutaline prior to allergen (P less than 0.05, Sign test). The data support the concept of in vivo inhibition of the mast cell mediator release reaction in atopic skin by terbutaline.

Adrenergic beta-Agonists↗

PubMed1986

Specific diagnosis of exogenous bronchial asthma in children.

Twenty-six children with exogenous asthma were studied by case history (CH), skin tests (ST), allergen-specific serum IgE (RAST), basophil histamine release (HR), and bronchial provocation tests (BPT). Nine standardized allergens were used from the following groups: house dust mite, animal dander, pollen, and mould fungi. For each allergen, the same batch was used throughout for all in vivo and in vitro studies, and the tests were carried out in a controlled, double-blinded manner, independent of each other. Different allergen concentrations were used and the results graded on a semi-quantitative scale. Approximately 120 comparable investigations, using in vivo and in vitro tests were carried out, followed by calculations of the sensitivity and specificity for each test alone, as well as for various combinations of tests, at various allergen-concentrations, and compared with the result of BPT, i.e. the true diagnosis. In this way the results could be used to directly compare the diagnostic value of each test, and combinations of tests. In general, a careful CH combined with ST gave the most reliable results. RAST was the most unreliable single analysis. When pollen or mite allergy was suspected, a combination of ST with either RAST or HR gave a further diagnostic reliability. HR as a single analysis did not seem to offer advantages over existing methods. None of the tests, or combinations of tests offered 100% sensitivity and specificity. By altering the concentration of allergen, the sensitivity, or specificity could be increased, but only at the expense of a decrease in the other. BPT is still necessary in many instances when a specific diagnosis is required.

Adolescent↗

PubMed1986

Influence of bacterial endotoxins on basophil histamine release. Potentiation of antigen- and bacteria-induced histamine release.

The histamine-releasing capability of lipopolysaccharides (LPS) was examined in human leukocyte suspensions. LPS alone did not release histamine, but was found to enhance the histamine release caused by anti-IgE. Also the IgE-mediated histamine release caused by specific antigens (allergens or bacteria) in sensitized individuals was enhanced by LPS. The potentiating effect of LPS was observed in grass pollen and dog dander allergic patients as well as in patients sensitized to E. coli or Staph. aureus bacteria. No potentiation was obtained by exposure to unspecific allergens or bacteria to which the persons were not sensitized. Bacteria can release histamine by immunological or nonimmunological mechanisms, and only the immunological histamine release was found to be potentiated by LPS. It is speculated that endotoxins reinforce release of histamine caused by allergens in allergic patients or by bacteria in persons sensitized to these microorganisms.

Allergens↗

PubMed1986

Long-term changes in inhalant allergy in asthmatic children.

In a representative cohort of 55 asthmatic schoolchildren the progress of the allergy per se was followed up prospectively for 8 years. Judged by clinical data, skin prick tests and RAST, a large majority of the children retained their allergies to pollen and animal danders. These allergens were predominant, whereas allergy to mites and moulds was less frequent. Serum IgE levels showed a strong tendency to remain high. Positive RAST and skin prick tests were also found in a substantial number of children with normal serum IgE concentrations.

Adolescent↗

PubMed1986

Histamine-dependent allergy blood test. Clinical evaluation.

Allergen-mediated histamine release from human leukocytes represents an important model for in vitro studies of allergic reactions. The purpose of this study was to determine whether the measurement of histamine released in allergic patients by radioenzymatic assay following mixing of their blood with common allergens represents a reliable index for diagnosis of atopic allergy. Three categories of allergens were used: 1) house dust and mite; 2) cat and dog dander; 3) trees, grasses and ragweed mixture. The presence of allergy was established by clinical history and intradermal skin testing in the study group of 150 patients. A significant allergen-mediated histamine release ranging from 4 to 65% of the total blood histamine content was observed in 96% of the patients with skin test sensitivity of greater than or equal to 3+. There was a significant correlation between skin testing and histamine release in terms of the allergens causing the response. Thus, the measurement of histamine by radioenzymatic technique following its release in blood in response to allergen challenge represents a clinically useful in vitro test for the diagnosis of atopic disease.

Allergens↗

PubMed1988

Passive sensitization of basophil leukocytes from non-atopic adult by plasma from allergic children.

Basophil leukocytes from a non-atopic donor, who responded well to anti-IgE, were depleted of their native membrane-bound IgE by acid treatment and passively sensitized with plasma containing either Phleum pratense-, Dermatophagoides pteronyssinus- or dog dander- specific IgE obtained from 18 allergic children. Histamine release was then performed on the passively sensitized cells and the results were compared with those of bronchial provocation test (BPT), allergen-specific serum IgE (RAST), skin prick test (SPT), and conventional histamine release test (HR). A high coincidence rate was found between BPT, RAST and histamine release after passive sensitization (HR-PS), but compared to HR, it was lower. This could be because several of the patients had non-responding basophils (i.e. no release after challenge with anti-IgE) in the conventional histamine release assay. The lower rate was not related to a lack of antigen-specific IgE, since after passive sensitization of basophils, anti-IgE and allergen provocation could induce release. It is concluded that plasma from allergic children with non-responding basophil leukocytes contain antigen-specific IgE capable of binding to Fc-receptors on the basophils of a non-atopic donor. In addition it was found that the plasma could change the cell reactivity (maximal histamine release) of the donor cells, since the amount of histamine released varied according to the plasma used for passive sensitization. The lack of histamine release response in some patients could be because their own membrane-bound IgE is unable to induce mediator release or, more likely, activation of one or more of the subcellular steps involved in the release is impaired.

Adolescent↗

PubMed1989

Influenza A virus enhances allergic histamine release. Effect of neuraminidase.

Histamine release was examined in leukocyte suspensions from patients allergic to house dust mite, grass pollen, birch pollen or cat dander. Influenza A virus was found to enhance the antigen-induced mediator release, but did not cause release of histamine from the cells per se. Also histamine release induced by anti-IgE in cell suspensions from normal individuals was enhanced by virus. The potentiating effect of influenza A virus might be due to neuraminidase on the surface of virus, since a similar effect was caused by a purified neuraminidase obtained from Vibrio cholerae, and the effect of virus as well as the neuraminidase was completely abolished by a potent neuraminidase inhibitor. The synergistic enhancement in IgE-mediated histamine release by virus could be of significance for the conversion from latent to manifest asthma.

Adult↗