Time course of the inflammatory response to histamine and allergen skin prick test in guinea-pigs

Plasma exudation and vasodilatation are key microvascular features of acute inflammation. Exudation and vasodilatation responses in the weal area after skin prick testing with histamine are essentially completed within 30 min. There is evidence to suggest that vasodilatation lasts considerably longer after provocation with allergen, but there is no information on the duration of plasma exudation. The purpose of this study was to measure the time course of the microvascular inflammatory response in the skin after histamine and allergen provocation. Skin prick tests were performed with histamine, allergen (ovalbumin) or saline (control) on guinea-pigs which were shaved on their backs. Radioactive ^113mIn was used to label transferrin as a plasma tracer. Radioactivity was recorded from the superficial part of the skin by external detection of conversion electrons from the decay of ^113mIn. The increase in count rate, corresponding to tracer accumulation by vasodilatation and/or plasma exudation, was used as a measure of the microvascular inflammatory response to skin prick test. The microvascular response was studied immediately and up to 30 min after provocation. The largest response to histamine and allergen occurred immediately after provocation. The exudative response then gradually declined to be absent after 25–30 min. Skin prick test with saline resulted in a small response of shorter duration. We conclude that the microvascular reaction to histamine as well as allergen provocation in guinea-pig skin has a rapid onset and a duration of ≈30 min.     

Dose response relationship of allergen, histamine, and histamine releasers in skin prick test and precision of the skin prick test method

The aim of the present investigation was to study the dose response relationship of allergen, histamine and histamine releasers in skin prick test (SPT) and the precision of the SPT method. In one experiment timothy allergen, histamine HCl, rabbit anti-human-IgE and compound 48/80 were studied in seven patients. In a second experiment timothy allergen and histamine and dog allergen and histamine were investigated in two groups of 10 patients. Histamine HCl 1 and 10 mg/ml induced weals about 15 and 25 mm2 (4.5 and 5.5 mm in diameter), respectively. The precision as expressed by the coefficient of variation was about 25% for histamine and 40% for allergen for weal areas greater than 10 mm2. Calculations of the regression lines to test the dose response relationships were based on the method of least squares. The best fit was to a log/log model. The slopes of allergen, histamine and histamine releasers were essentially parallel within patients. The median slope of allergen was estimated to about 0.4 based on weal areas and 0.2 based on mean weal diameters. Furthermore, no significant differences were found between the lower and upper parts of the dose response curves of allergen and histamine, although there was a tendency towards steeper slopes at lower concentrations. These results show that histamine concentrations greater than or equal to 1 mg/ml should be used as positive control in SPT and that histamine releasers do not offer advantages over histamine as reference substances in SPT. A common slope for the dose response relationship of allergen and histamine can be used for the estimation of skin sensitivity.     

Dose response curve of allergen and histamine in skin prick tests

Twenty-six Dermatophagoides pteronyssinus (D.pt.) sensitive subjects were skin prick tested in duplicate with 15 concentrations of D.pt. ranging from 0.0018 to 17.8 mg/ml, 15 concentrations of the major allergen of D.pt., antigen P1, ranging from 0.0002 to 1.88 mg/ml and 15 concentrations of histamine dihydrochloride solution ranging from 0.048 to 114.0 mg/ml. Weal areas and concentrations were transformed by taking logs and linear and non-linear regression curves fitted, allowing for confounding variables, such as subject, and interactions. The weal areas over all concentrations fitted "S" shaped curves with essentially straight central portions, parallel between materials, with differences between subjects but parallel within subjects. The dose response curves of P1 and D.pt. were coincident when the concentrations were adjusted to allow for differences in potency. The concentrations of allergen and histamine commonly used for standardisation purposes will give weals that can be plotted along a straight line, but at higher and lower concentrations the response will tail off. This accounts for previously ambiguous results. Standardisation of allergens using 10 mg/ml histamine is preferable to 1 mg/mg.     

Standardization of food allergen extracts for skin prick test.

The aim of the study was to standardize and evaluate technically optimized food allergen extracts for use in skin prick test (SPT). The standardization procedure comprised 36 allergic histories in 32 food allergic patients with 21 healthy, non-atopic individuals serving as controls. The patients had a history of allergic symptoms upon ingestion of either cow's milk (n=3), hen's egg (n=9), wheat (n=4), hazelnut (n=14) or cod (n=6). They also had specific IgE in serum to the food in question and a positive SPT with a fresh preparation of the food. The diagnosis had been confirmed by a double-blind, placebo-controlled food challenge, except for the hazelnut-allergic patients. The controls were subjected to an open food challenge with all the foods to ensure tolerance. The standardization was performed by means of titrated SPT in accordance with the guidelines on biological standardization from the Nordic Council on Medicine. Regression analysis of the skin wheal areas was performed for each patient and the median protein concentration of allergen preparation (median Ch10) eliciting a wheal area of the same size as histamine 10 mg/ml was calculated. The median Ch10 was 0.56 mg/ml for milk, 0.88 mg/ml for egg, 5.4 mg/ml for wheat, 2.1 mg/ml for hazelnut and 0.017 mg/ml for the cod extract. The sensitivity of the median Ch10 estimated from the SPT data was 1 for milk, 0.98 for egg, 1 for wheat, 1 for hazelnut and 0.87 for the cod extract. The allergenic activity of the hazelnut extract was further investigated by leukocyte histamine release (HR) and immunoblotting experiments using sera from 27 hazelnut allergic patients. The clinical sensitivity of the optimized hazelnut extract evaluated by HR was 0.78 compared to 0.30 for a commercially available hazelnut extract (Soluprick). Immunoblotting results showed a stronger IgE binding capacity and additional IgE-binding bands of the optimized hazelnut extract compared with the Soluprick extract.     

几种户尘螨过敏原点刺液的临床评价

评价户尘螨过敏原点刺皮试液用于诊断和筛查尘螨过敏的意义。方法：对219例患者进行户尘螨过敏原点刺试验和户尘螨sIgE检测。点刺试验所用点刺液为本课题组研制的点刺液（A 组）、ALK公司的户尘螨点刺液（B组）和ALK公司的混合螨（户尘螨和粉尘螨）点刺液（C组）,以sIgE为诊断标准。统计分析采用SPSS 11．0软件,对三组户尘螨过敏原点刺试验进行ROC曲线评价。结果：A、B和C组风团平均直径中位数为0.43cm、0.35cm和 0.28cm。所有入选者点刺试验后1h,A组有5例点刺试验后出现局部反应,2h左右自行缓解;有2例出现迟发反应,需1天左右才消退;B组和C组各有3例点刺试验后出现局部反应,B组和C组各有2例有迟发反应。所有患者均无局部皮疹、全身性不良反应。三组户尘螨过敏原点刺试验的ROC曲线下面积分别为0.765、0.801、0.782,说明户尘螨过敏原点刺皮试液对诊断户尘螨过敏具有较高准确性。以户尘螨sIgE为标准的点刺试验灵敏度和特异度：A、B 和C组点刺液的灵敏度分别为92.4％、87.0％和81.5％,特异度分别为60.6%、73.2%和74.8%。结论：户尘螨点刺液可作为用于尘螨过敏的诊断试剂,有良好的灵敏度和特异度,具有临床筛查和诊断意义。     

Use of standardized and conventional allergen extracts in prick skin testing.

This study examined whether commercially available conventional and standardized allergen extracts differ enough in potency to affect routine prick skin test results. Extracts of white oak, timothy, Bermuda, Russian thistle, short ragweed, sagebrush, Alternaria, and cat dander were examined in allergic patients and in nonatopic subjects with no personal or family history of asthma, rhinitis, or eczema. Conventional nonstandardized extracts (1:10 or 1:20 wt/vol) from two sources were compared with three concentrations (100,000, 10,000, and 1000 AU/ml) of a single standardized extract. Preparations were compared in the allergic patients with computerized planimetry, and in all patients and subjects with a conventional skin test grading system. Skin test area for the conventional extracts generally fell between the 10,000 and 100,000 AU/ml concentrations of the standardized extract. Skin test reactivity to at least one allergen extract occurred in 31% of the nonatopic subjects; there was no difference between the number of 3+ and 4+ reactions for conventional and standardized extracts. Results indicate that standardized and conventional extracts are frequently similar, but are not directly interchangeable.     

Reproducibility of the skin prick test

Reproducibility of the skin prick method of testing for allergy was studied in 20 subjects examined by four nurses. Hypodermic needles were used for pricking and the test panel included a histamine control, a diluent control, and nine allergens. The reproducibility of the method was best when the size of the weal reaction caused by an allergen was expressed as the geometric area of the weal. When the weal reaction was expressed as the ratio of the weal reaction caused by an allergen to that caused by histamine, the reproducibility of the method was decreased considerably. When the ratios were further classified into three class ratings, reproducibility was very low. The reduction in reproducibility was due to the low reproducibility of histamine reactions. According to these results, at least in epidemiological studies the weal reactions should be expressed as geometric areas. In clinical practice it might also be preferable to express prick test results as the diameters of the weals without adjusting them by histamine reactions.     

Inhibition of different dosages of oxatomide or placebo on skin prick test and nasal allergen provocation.

In this two-stage, double-blind study, we evaluated the effects of different dosages of oxatomide (1 and 2 mg/kg/day) on nasal provocation and skin reaction wheal induced by grass-pollen challenge. Children with a positive history of allergic rhinoconjunctivitis and positive responses to skin prick test and nasal provocation test to grass pollen were studied out of season. The results obtained with 1 mg/kg/day of oxatomide demonstrated no significant difference in wheal areas and nasal secretion induced by allergen challenge between treated and untreated patients. The administration of 2 mg/kg/day demonstrated a significant suppression in wheal reaction and nasal secretion induced by specific challenge.     

Egg and milk allergy in adults: comparison between fresh foods and commercial allergen extracts in skin prick test and histamine release from basophils

The ability of skin prick test (SPT) and histamine release from basophils (HR) to diagnose clinical type I allergy to egg and milk was investigated as compared with double-blind, placebo-controlled food challenge (DBPCFC) in 17 adults suspected of type I egg and/or milk allergy. In both SPT and HR, commercial allergen extracts commonly used for SPT were compared with fresh, standardized foods. With commercial extracts the overall sensitivities of SPT and HR were 0.75 and 0.56 respectively, and none of the tests showed concordance with DBPCFC. With fresh, standardized foods the overall sensitivities of SPT and HR were 1.00 and 0.89 respectively, and both tests now showed a significant concordance with DBPCFC (P < 0.05). Specificity was only slightly improved in SPT, and unchanged in HR. Thus, the use of fresh, standardized foods significantly improved the outcome of both tests, as regards to sensitivity and concordance with DBPCFC. The diagnostic ability of SPT and HR appear to be strongly influenced by the allergen quality.     

Studies on the osmophilic fungus Wallemia sebi as an allergen evaluated by skin prick test and radioallergosorbent test

Recently, Wallemia sebi, a species of osmophilic fungi, has been abundantly detected in house dust using low water activity media. In this study, allergenic activity of W. sebi was assessed by skin prick test and radioallergosorbent test (RAST) in 74 asthmatic patients ranging from 6 to 32 years of age. Aspergillus fumigatus and house dust were used for comparison. In skin prick test, W. sebi extract, A. fumigatus extract and house dust extract elicited positive reactions in 4 (5.4%), 4 (5.4%) and 51 (68.9%) patients, respectively. RAST showed positive results in 14 subjects (18.9%) for W. sebi extract, in 8 (10.8%) for A. fumigatus extract and in 59 (79.7%) for house dust extract. These results indicated that some asthmatic individuals showed immediate-type hypersensitivity to W. sebi, and which means this fungal species may be of importance to atopic diseases as a causative agent.     

The links between allergen skin test sensitivity, airway responsiveness and airway response to allergen

BACKGROUND: The allergen-induced early asthmatic response [provocation concentration (PC)20, the concentration causing a 20% forced expiratory volume in 1 s (FEV)1 fall] depends on the level of IgE sensitivity and the degree of nonallergic airway hyperresponsiveness (AHR) and can be predicted from histamine PC20 and allergen skin test endpoint. OBJECTIVES: We examined the relationships between allergen PC20, methacholine PC20, and allergen skin test endpoint and assessed the accuracy of both the histamine PC20-based prediction of allergen PC20 (using methacholine) and a new methacholine PC20-based prediction equation. METHODS: From 158 allergen challenges, the allergen PC20, the methacholine PC20, and the skin test endpoint were recorded and relationships between these three were sought. We compared the measured allergen PC20 to that predicted from the previous histamine PC20-based and the new methacholine-based formulae. RESULTS: In single regressions, allergen PC20 correlated with both methacholine PC20 (r=0.25, P=0.0015) and skin test endpoint (r=0.52, P <0.00005). The relationship was improved by multiple regression of log-allergen PC20vs. log-methacholine PC20 and log-endpoint (r=0.61, P <0.00005). The histamine-based formula predicted allergen PC20 to within 2 doubling concentrations in 80% and within 3 in 92%. The new methacholine-based formula to within 2 and 3 concentrations in 81% and 94%, respectively; only nine of 158 subjects were outside the 3 concentrations. CONCLUSIONS: We have confirmed the dependence of the allergen-induced early asthmatic response upon the level of allergic sensitivity and the degree of AHR, the latter as assessed by methacholine challenge. The allergen PC20 can be predicted to within 3 doubling concentrations in 94% of cases.     

Association between alcohol consumption and skin prick test reactivity to aeroallergens.

BACKGROUND: A few studies have indicated a positive association between consumption of alcohol and allergic sensitization in age and socioeconomically heterogeneous populations. OBJECTIVE: To investigate the association between consumption of alcohol and allergic sensitization in a young homogenous population of high social class (a group with a suspected high prevalence of sensitization). METHODS: A total of 1,668 students aged 18 to 35 years recruited from universities in Copenhagen, Denmark, underwent skin prick testing (SPT) in October or November 2002 and completed a questionnaire about respiratory disease and lifestyle habits, including alcohol consumption. SPT positivity was defined as a positive reaction (> or =3 mm) against at least 1 of 10 common inhalant allergens. RESULTS: Before and after adjustment for sex, age, smoking, atopic predisposition, and pet keeping, no significant association was found between alcohol consumption (including type of beverage) and SPT positivity. Increasing alcohol consumption was significantly negatively associated with asthma symptoms and hay fever symptoms. CONCLUSIONS: Alcohol consumption does not favor SPT positivity, but cumulated effects were not addressed in the present study. Individuals with asthma or hay fever symptoms seem to reduce alcohol intake (a healthy drinkers' effect).     

Basophil histamine release and airway response to mite allergen in atopic dermatitis.

Twelve patients with atopic dermatitis (AD) were subjected to in vitro histamine release from peripheral blood leukocytes (basophils) and in vivo bronchial inhalation challenge using house dust mite (Dermatophagoides farinae) allergen. Not only seven patients with asthmatic history but also five patients without asthma responded to both the in vitro and the in vivo challenges. A significant correlation was observed between HR30 (a mite concentration producing a 30% release of total cellular histamine) and PC20 allergen (a mite concentration producing a 20% fall in FEV1). There was also a significant correlation between MHR (maximal histamine release) and the maximal fall in FEV1. The relationship held for both AD patients with asthma and without asthma. These results suggest that histamine release induced by the house dust mite allergen is a good in vitro test for predicting the bronchial response to this allergen. They also suggest that these tests are not disease specific, but are valuable in evaluating the degree of atopic state in a subject.     

Longitudinal variability of skin prick test results

The skin prick test (SPT) is a commonly used procedure for assessing a specific sensitization. The longitudinal variability of test results is of interest for clinical as well as epidemiological investigations. The sensitization to four common aeroallergens (grass pollen, birch pollen, Dermatophagoides pteronyssinus, cat dander) is investigated within the framework of three consecutive SPTs at 11-month intervals for a population of 587 schoolchildren. The prevalence of sensitization based on a weal diameter of at least 2 mm was between 12.9% (cat dander) and 23.9% (grass pollen) in the initial testing. The positive predictive values of the initial SPT were between 75.3% (birch pollen) and 88.2% (cat dander) for the two subsequent SPTs. In the case of initially negative tests with positive second and third SPTs the incidence ranged between 3.2% (cat dander) and 4.3% (birch pollen) per year. A clear increase in the intensity of reaction in subsequent tests was observed in a number of probands testing positively in the initial SPT. In conclusion, our data indicate a high long-term stability of a specific sensitization to aeroallergens in SPT.     

Clinical application of histamine prick test for food challenge in atopic dermatitis.

Determining positive food challenges are not easy as there is an absence of simple and objective tests. Histamine, an essential mediator for allergic reactions, is involved in the pathogenesis of atopic dermatitis (AD) and food challenges can change histamine levels. The significances of a prick test with histamine (histamine prick test, HPT) relating to the interpretation of food challenges in AD were evaluated. A total of 467 AD patients participated in this study. Skin prick tests, identification of specific IgE and open food challenge were conducted for the identification of food allergy. Elimination diet was performed with HPT. HPTs were conducted before and after food challenges. The wheal sizes by HPT were significantly decreased after an elimination diet. The relative changes of wheal sizes significantly correlated with those of clinical severity scores in AD patients (p<0.001). The wheal sizes in HPT were increased with a positive provocation in open food challenges. In conclusion, HPT may be a simple and objective test to interprete the results of food challenges in patients with AD. The exact mechanisms of the changes in skin reactivity by HPT need further investigation.     

Topical dermal anaesthesia inhibits the flare but not the weal response to allergen and histamine in the skin-prick test

The effect of topical dermal anaesthesia on the immediate allergic skin reaction was evaluated in a double-blind, randomized, placebo-controlled study. Twenty-one patients with strictly seasonal allergic rhinitis, confirmed by a positive skin test for the respective pollen allergen, were studied in the pollen-free winter months. Skin-prick tests for one pollen allergen and histamine were performed after pre-treatment of the skin for 1 hr with an emulsion of lidocaine and prilocaine (EMLA®) and the equivalent vehicle on different test sites. The skin-prick tests were made with a preloaded standardized test needle (Phazet®). The area of the induced weal-and-flare reaction was measured and subsequently calculated with the help of a digitizer served by a microcomputer. The topical dermal anaesthesia induced a reduction of the flare response to histamine by 49% (P < 0.01) and allergen by 21% (P < 0.05). No reduction of the histamine- and allergen-induced weal response were noted. Our findings indicate that the treatment did not affect the allergen-induced release of inflammatory mediators and the vascular leakage induced by these mediators. However, this study seems to confirm earlier suggestions that the flare response is partly mediated through neural reflex activity as it was ameliorated by topical anaesthesia. Furthermore, from a clinical point of view, this study shows that it is possible to perform a valid skin test, without any associated itching and pain, if only the weal response is taken into account n i the evaluation of the skin-prick test.     

Diagnostic value of scratch-chamber test,skin prick test,histamine release and specific IgE in birch-allergic patients with oral allergy syndrome to apple

BACKGROUND: The aim of the study was to examine the diagnostic value of skin prick test (SPT), scratch-chamber test (SCT), histamine release (HR) and specific immunoglobulin E (IgE) in birch-allergic patients with oral allergy syndrome to apple. METHODS: Ten birch-allergic patients with oral allergy syndrome to apple and 10 control subjects were included. All were tested with SPT, SCT, HR and specific IgE [CAP, Pharmacia, Sweden and Magic Lite (ML), ALK-ABELLO, Denmark]. RESULTS: The SPT with apple, acetone extract of apple (A72) and commercial apple extract showed sensitivities of 0.80, 0.90 and 0.10, respectively. The SCT with the same extracts showed sensitivities of 0.30, 0.50 and 0.20, respectively. The sensitivity of specific IgE to apple were 0.90 (CAP) and 0.10 (ML). The sensitivity of the HR test was 90% (A72), and 25% using the commercial extract. CONCLUSION: The SPT and HR test with apple and A72 showed a good diagnostic value with a sensitivity of more than 70% and a specificity of 100%. The SCT showed a poor sensitivity to apple, A72 and commercial apple extract. The ML test was not suitable in detecting specific IgE to apple compared with the CAP test. In daily practice a detailed case history about symptoms of oral allergy syndrome combined with a SPT with fresh apple peel or A72 will be useful.     

Relationships of total IgE level, skin prick test response, and smoking habits

Epidemiologic observations on 331 men showed that increased serum IgE concentration was associated with a wheal response to skin prick testing, but also to an erythema response in the absence of any wheal, and to heavy smoking. The association between IgE and the various skin prick test responses remained after taking into account smoking and asthma.