High burden of clonal mast cell disorders and hereditary α-tryptasemia in patients who need Hymenoptera venom immunotherapy.

BACKGROUND: In patients who require venom immunotherapy (VIT), there is a need to identify underlying mast cell (MC) disorders since these may affect the risk and severity of future sting reactions and the long-term effectiveness of VIT. METHODS: 1319 individuals with Hymenoptera venom allergy (HVA) who needed VIT from referral centers in Slovenia, Austria, Croatia, and Poland underwent examination for KIT p.D816V in peripheral blood leukocytes (PBL) using a highly sensitive PCR test and tryptase genotyping by digital droplet PCR. We also included 183 control individuals with large local reactions (LLRs) to Hymenoptera stings and with asymptomatic sensitization to Hymenoptera venoms. RESULTS: 285 of 1319 individuals recommended for VIT (21.6%) were positive for KIT p.D816V in PBL, preferably those who present with severe reaction (33.9% [n = 207 of 610] with Ring-Messmer grade 3-4 vs. 11% [n = 78 of 709] with Grade 1-2; p < .0001), whereas only 1.3% (n = 2 of 152) of controls with LLR and none with asymptomatic sensitization (n = 31) had KIT p.D816V. KIT p.D816V allelic burden was higher in those with severe reaction (median 0.018% [n = 207] in Grade 3-4 vs. 0.001% [n = 78] in Grade 1-2; p < .0001), and the majority had normal baseline serum tryptase levels (69% [n = 196 of 285]). All KIT p.D816V-positive individuals (n = 41) who underwent bone marrow (BM) biopsy were found to have underlying clonal diseases, principally BM mastocytosis. HαT was also associated with severe HVA and symptoms (p < .01), and remarkably, 31.0% (n = 31 of 100) were found to have concomitant KIT p.D816V. Concomitant HαT and KIT p.D816V showed an additive effect, and having both was associated with the highest risk for severe HVA, even higher than having either HαT or KIT p.D816V alone (OR = 3.8; p < .01). CONCLUSIONS: By employing prospective universal tryptase genotyping and examination for KIT p.D816V in PBL in large HVA populations, we have demonstrated a high burden of clonal MC disorders and HαT in patients who require VIT.

Optimization of Basophil Activation Test in the Diagnosis and Qualification for Allergen-Specific Immunotherapy in Children with Respiratory Allergy to the House Dust Mite Dermatophagoides pteronyssinus.

The aim of this study was to optimize a basophil activation test in the detection of allergy to the house dust mite Dermatophagoides pteronyssinus in children with allergic respiratory diseases. This study involved 32 cases, 13 girls and 19 boys aged 4-17 years, with perennial asthma or allergic rhinitis caused by D. pteronyssinus. The control group consisted of 13 girls and 19 boys aged 4-17 years with seasonal allergic asthma or rhinitis provoked by Timothy or birch pollen. House dust mite (HDM) allergy was excluded in the controls based on their medical history, skin prick test (SPT) results and sIgE determination. In all patients, a basophil activation test (BAT) was performed with five dilutions of D. pteronyssinus allergen (the dilution series ranged from 22.5 to 0.00225 ng/mL). The results were analyzed by using the receiver operating characteristics (ROC) to determine the optimal allergen concentrations, outcome measures and cut-off points that would differentiate most accurately between HDM-allergic and non-allergic patients. As a "gold standard", criteria for allergen-specific immunotherapy with D. pteronyssinus or respective pollens were applied by an experienced pediatric allergist following the guidelines of the European Academy of Allergy and Clinical Immunology. The highest diagnostic efficiency was yielded by the protocol assuming a cut-off value of 9.76% activated basophils after activation with a single allergen concentration of 2.25 ng/mL (sensitivity 90.6%, specificity 100%). This protocol yielded 3 (4.7%) misclassifications, all false negative, when compared with the "gold standard". There was a strong correlation with the BAT results at 22.5, 2.25 and 0.225 ng/mL (respectively r = 0.90 and r = 0.78, p < 0.001), as well as between the BAT at 2.25 ng/mL and SPT (r = 0.82, p < 0.001) and between the SPT and sIgE levels (r = 0.78, p < 0.001). High cross-reactivity between D. pteronyssinus and D. farinae was confirmed based on the BAT at 22.5 ng/mL (r = 0.82, p < 0.001). In conclusion, the BAT showed very good concordance with the result of a meticulous process of decision-making that combined validated allergy tests (SPT, sIgE) with expert guidelines, specialist knowledge and experience. Facing the risk of the incorrect qualification of patients for costly, long-lasting and potentially risky allergen-specific immunotherapy, the inclusion of a basophil activation test into diagnostic process seems fully justified.

Non-IgE Mediated Hypersensitivity to Food Products or Food Intolerance-Problems of Appropriate Diagnostics.

The diagnosis of food intolerance is tricky due to the different etiologies of adverse reactions. There is also a lack of clear rules for interpreting alternative tests used to diagnose these problems. The analyses of IgG4 concentration in serum or cytotoxic tests became the basis of elimination diets. However, it can result in nutritional deficiencies and loss of tolerance to eliminated foods. Our study aimed to assess the necessity of food elimination in four cases with food intolerance symptoms based on alternative diagnostic tests. Four patients without food allergies, who manifested diverse clinical symptoms after food, were presented due to the following factors: clinical history, diagnostic tests, elimination diet, and filaggrin gene (FLG) mutation. It was found that higher IgG4 levels against foods and higher cytotoxic test values are not clinically relevant in each of the studied individuals. They should not be decisive for the elimination of food products. The study of FLG-SNVs revealed the association of some clinical symptoms in patients with hypersensitivity to several food allergens and reported genetic variants in the FLG gene.

Genetic variants of SERPING1 gene in Polish patients with hereditary angioedema due to C1 inhibitor deficiency.

Hereditary angioedema due to C1-inhibitor deficiency (C1-INH-HAE) type I and II is a rare and life-threatening disease caused by SERPING1 gene mutations. Previous genetic studies indicated a wide spectrum of disease-associated variants in the SERPING1 gene and often lack of correlation with patient's phenotypes. The aim of this study was to evaluate the presence, type, and localization of mutations in the SERPING1 gene in 41 Polish patients with C1-INH-HAE and their relation with case/family history, type of C1-INH-HAE, fC1-INH, age of onset, and disease severity. Sanger sequencing and MLPA method were used for detection of disease-associated variants. In 34 (82.9%) patients, mutations located in various regions of SERPING1 gene were revealed. The detected alterations in patients with C1-INH-HAE type I differed and were positioned in various exons/introns of the SERPING1 gene. The most frequent disease-associated variants appeared in exon 3 (especially in type I) and in exon 8 (type I and II). Out of 20 different disease-causing variants, 9 were not previously described. We did not find any relation between the type and location of the mutations and no type of features included in phenotype evaluation of the patients, such as case and family history, type of C1-INH-HAE, age of onset, biochemical parameters, or severity of disease.

Prevalence and potential risk factors of urticaria in the Polish population of children and adolescents.

INTRODUCTION: Urticaria is a frequent, mast cell driven disease of great social significance as it may lead to disablement, impair quality of life and affect performance at work and school. AIM: A study on the prevalence and potential risk factors for acute and chronic urticaria in the population of children and adolescents living in Krakow, one of the major cities in Poland. MATERIAL AND METHODS: In this paper we present a study on the prevalence and potential risk factors of acute and chronic urticaria in the population of children aged 7-8 years old and adolescents aged 16-17 years old living in Krakow, one of the major cities in Poland. The relation of urticaria prevalence and exposure to polluted air was also investigated. In the period of 3 years between 2016 and 2018 we enrolled 17082 participants (9287 children and 7795 adolescents). RESULTS: Lifetime occurrence of at least one episode of urticaria was reported by 3.3% of respondents (3.6% in the children group and 2.8% in adolescents). The most frequently mentioned causative agent was medicines in children and food in adolescents. Chronic urticaria occurred in 2.1% of children and 1.1% of adolescents. We found no relationship between the volume of traffic and the distance of residence from the high-traffic street to the frequency of urticaria. CONCLUSIONS: A higher incidence of urticaria in the younger age group may suggest an upward trend in the incidence of this disease in the near future. The incidence of chronic urticaria in children turned out to be higher than in the European population. As our research results show, the incidence of urticaria in children and adolescents does not depend on exposure to polluted air.

Component-resolved diagnosis (CRD) of food anaphylaxis

Anaphylaxis is an increasing problem in public health. The food allergens (mainly milk, eggs, and peanuts) are the most frequent cause of anaphylaxis in children and youth. In order to define the cause of anaphylaxis, skin tests, the determination of the concentration of specific IgE in the blood and basophil activation test are conducted. In vitro tests are preferred due to the risk of allergic response during in vivo tests. Component-resolved diagnosis (CRD) is an additional tool in allergology, recommended in the third level of diagnostics when there are diagnostic doubts after the above mentioned tests have been carried out. The paper presents 3 cases of patients with anaphylactic response, and the application of CRD in these patients helped in planning the treatment. Patient 1 is a 4-year-old boy with diagnosed atopic dermatitis and bronchial asthma reported an anaphylactic shock at the age of seven months caused by cow's milk and the exacerbation of bronchial asthma after eating some fruit. Patient 2 is a 35-year-old woman who has had anaphylactic shock three times: in June 2015, 2016, and 2017 and associates these episodes with the consumption of dumplings with a caramel, bun, and the last episode took place during physical exertion few hours after eating waffle. Patient 3 is a 26-year-old man with one-time loss of consciousness after eating mixed nuts and drinking beer. CRD offers the possibility to conduct a detailed diagnostic evaluation of patients with a history of anaphylactic reaction.

The usefulness of the basophil activation test in monitoring specific immunotherapy with house dust mite allergens.

INTRODUCTION: In clinical practice, reliable tools for monitoring specific immunotherapy (SIT) are of utmost importance. AIM: To assess the usefulness of the basophil activation test (BAT) in monitoring SIT in paediatric patients with allergy to house dust mites (HDM). MATERIAL AND METHODS: Thirty-one children qualified for SIT with HDM, of whom 21 completed the SIT during the observation period. The BAT was carried out prior to commencing the SIT (time point BAT1) and upon finishing the initial pack of allergy vaccine (cumulative dose of allergen 12487.5 PNU; BAT2), as well as after the second vaccine pack (cumulative dose of allergen 23750.0 PNU; BAT3). Peripheral blood of the patients was stimulated with allergen solutions in five concentrations from 0.00225 ng/ml to 22.5 ng/ml. Basophil activation was measured by CD63 expression in flow cytometry. RESULTS: For the allergen concentration of 0.225 ng/ml, a statistically significant decrease in median basophil activation was observed, from 51.29% at BAT1 to 8.48% at BAT2 (p = 0.004) and 4.21% at BAT3 (p < 0.001). For the allergen concentration of 0.0225 ng/ml, a statistically significant decrease was seen between BAT1 (1.72%) and BAT3 (0.21%, p = 0.01). Median CD-sens index decreased significantly from 1099.02 at BAT1 to 179.31 at BAT2 (p < 0.002) and 168.04 at BAT3 (p < 0.001). CONCLUSIONS: There is a significant decrease in BAT results in the course of specific immunotherapy with HDM allergens in children, with the optimum allergen concentration for monitoring basophil response at 0.225 ng/ml. The CD-sens index seems to be a better monitoring parameter than the plain percentage of CD63-expressing basophils.

Simplification of intradermal skin testing in Hymenoptera venom allergic children.

BACKGROUND: The direct comparison between children and adults with Hymenoptera venom anaphylaxis (HVA) has never been extensively reported. Severe HVA with IgE-documented mechanism is the recommendation for venom immunotherapy, regardless of age. OBJECTIVE: To determine the differences in the basic diagnostic profile between children and adults with severe HVA and its practical implications. METHODS: We reviewed the medical records of 91 children and 121 adults. RESULTS: Bee venom allergy was exposure dependent, regardless of age (P < .001). Atopy was more common in children (P = .01), whereas cardiovascular comorbidities were present almost exclusively in adults (P = .001). In the bee venom allergic group, specific IgE levels were significantly higher in children (29.5 kUA/L; interquartile range, 11.30-66.30 kUA/L) compared with adults (5.10 kUA/L; interquartile range, 2.03-8.30 kUA/L) (P < .001). Specific IgE levels for culprit insect venom were higher in bee venom allergic children compared with the wasp venom allergic children (P < .001). In adults, intradermal tests revealed higher sensitivity, accompanied by larger area of skin reactions, regardless of type of venom. At concentrations lower than 0.1 µg/mL, 16% of wasp venom allergic children and 39% of bee venom allergic children had positive intradermal test results. The median tryptase level was significantly higher in adults than in children for the entire study group (P = .002), as well as in bee (P = .002) and wasp venom allergic groups (P = .049). CONCLUSION: The basic diagnostic profile in severe HVA reactors is age dependent. Lower skin test reactivity to culprit venom in children may have practical application in starting the intradermal test procedure with higher venom concentrations.

[Recent advances in the in vitro diagnosis of drug allergy].

Allergic reactions to drugs are a matter of great concern, both for patients and health care professionals. The diagnosis is primarily based on a clinical history and in vivo tests, but they have some significant limitations in a range of clinical situations. This review presents currently available in vitro methods for the identification of the culprit drug in drug-induced immediate reactions (specific IgE determination, basophil activation test), T-cell-mediated drug allergy (lymphocyte transformation test, enzyme-linked immunosorbent spot assay, cell activation markers and cytokine release) and in nonallergic hypersensitivity to nonsteroidal anti-inflammatory drugs. Perspectives for further improvement of the in vitro diagnosis of drug allergy are also discussed.

The effects of air pollution on the development of allergic diseases.

During the past years, air pollution created by humans has become a major problem increasing the public awareness. The incidence of allergic disease is high and continues to increase in populations of urban areas of Westernized countries throughout the world. The aim of this article is to summarize current knowledge of the effects of air pollution on the development of allergic diseases. In the paper the effect of outdoor and indoor air pollution is described with potential and proven mechanisms of action on the development of allergic diseases.

[Acquired angioedema ­ clinical characteristic of the patients diagnosed in 2012-2016 with acquired C1 inhibitor deficiency].

Acquired angioedema is a rare disease caused by a deficiency of C1 esterase inhibitor with recurrent swelling symptoms. It may occur in the course of lymphoproliferative disorders or autoimmune diseases. Symptoms resemble hereditary angioedema, and the only differentiating features is negative family history, late onset of symptoms and accompanying lymphoproliferative disorder. The aim of the study was to analyze the cases of acquired angioedema. The retrospective analysis of 341 patients from the registry of patients with C1 inhibitor deficiency. Results: We identified 4 patients among 119 with HAE (3.57%) diagnosed in this same period of time 2012-2016 who fulfilled the criteria of acquired edema. In two cases the primary reason of angioedema was lymphoproliferive disease, in two monoclonal gammapathy of unknown reason. We analyzed also the results of laboratory tests C4, C1 inhibitor, C1q. In all cases the face was dominated localization. After the treatment of primary lymphoproliferive disease, in two cases, we observed total remission of angioedema. Only one patient with gammapathy require treatment with C1 inhibitor during the attacks. In these case we observed both plasma deriver, and recombinant C1 inhibitor were effective.

[Honey bee venom allergy diagnostics].

Honey bee venom (Apis mellifera) is a mixture of many components, both amines, peptides enzymes and toxins. Most of the enzymes are allergens with different allergenic potential. It may affect not only to the symptoms, it may also contribute to the success of immunotherapy. In some cases the problem of a bee venom allergy diagnosis is difficult due to the imperfection of the classical diagnostic methods. In the present work reports in the literature regarding the bee venom allergy diagnosis on the basis of identification of the venom allergen sIgE to components are summarized. The use of recombinant allergens has allowed for the study of sIgE against the allergenic components, and thus determination of the individual profile of allergy. Extending the test panel of bee venom components to rApi m10 and rApi m3 compared to rApi m1 increases the diagnostic sensitivity by identifying new allergens that are not always present in the available preparations used in immunotherapy.

[The role of the basophil activation test (BAT) in qualification for specific immunotherapy with inhalant allergens].

Qualification for specific immunotherapy (SIT) according to the guidelines of the European Academy of Allergy and Clinical Immunology (EAACI) includes medical history, skin prik tests (SPT) and/or measuring the concentration of sIgE. It is necessary to perform additional diagnostic tests in case of discrepancies between the history and the results of SPT/sIgE or differences between SPT and sIgE. Basophil activation test (BAT) assesses the expression of activation markers of these cells, eg. CD63 and CD203c after stimulation. The aim of our study was to evaluate the usefulness of BAT in the qualification for the SIT in comparison to the SPT and sIgE and in case of discrepancies between the results of SPT and sIgE. The study included 30 patients with allergic rhinitis (AR) caused by allergy to house dust mite (Dermatophagoides pteronyssinus, Dp) or birch pollen qualified for SIT. All patients had SPT, sIgE and BAT determination. The group of patients with allergy to birch was a control group for Dp allergic and vice versa. BAT with CD63 antigen expression was performed using a Flow2CAST test. Basophils were stimulated with allergen preparation (50, 500, and 5000 SBU/ml concentrations). BAT results were expressed as a stimulation index (SI). For optimal concentrations of 50 and 500 SBU/ml parameters comparing BAT to SPT and sIgE as the gold standards were consecutively: sensitivity 82-100% and 93-100%, specificity 50-94% and 47-89%, positive predictive value 65- 94% and 61-87%, negative predictive value 86-100% and 93-100%. Correlation BAT - SPT and BAT - sIgE ranged within 0.59 to 0.84 and 0.51 to 0.72. BAT was helpful in 2 of 30 patients with incompatible results of SPT and sIgE. Optimal concentrations for basophil stimulation are 50 and 500 SBU/ ml. BAT may be useful diagnostic tool in the qualification for the SIT in case of discrepancies between the results of SPT and sIgE.

[Exposure to alder, hazel and birch pollen and PM10 dust concentration in Krakow in 2010-2015].

PM10, one of the air pollution components has a significant effect on plant pollen by the deposition on the surface of pollen grains, morphological changes of pollen walls and interference in the structure of the allergenic proteins. The aim of the study was to verify whether and in what weather conditions the concentration of pollen of alder, hazel and birch coincides with the elevated level of PM10 in Kraków, in 2010-2015. In the days when the alder pollen concentration exceeded 95 PG/m3, causing severe inhalant allergy symptoms and the concentration of hazel pollen was >80 PG/m3 and the admissible level of PM10 was exceeded, the maximum air temperature ranged from 5.1 to 22.6oC, the days were dry (humidity <60%), without precipitation and mostly windless. High concentration of birch pollen and exceeded admissible level of PM10 concentration were found in more than 60% of the days in the city center. In 70.3% of the days in which the birch pollen concentration reached values that caused the symptoms of asthma (>155 PG/m3), the dust concentration was exceeded (>50µg/m3), and rainfall and wind speed of 1-3 m/s occurred. Symptoms in people allergic to trees pollen in Kraków can be exacerbated during periods when the standard limits of PM10 are exceeded. Such situations have occurred mostly in March (pollen of alder and hazel) and April (birch pollen), especially in the city center.

[Diagnosis of food allergy]. / Diagnostyka alergii pokarmowej.

Food allergy is most often linked to the type I allergic reaction, while IgE-dependent mechanism causes symptoms in only about 50% of patients. If symptoms are coming from other types of allergic reactions we do not have enough standardized diagnostic methods. The purpose of our review is to discuss the possibilities of diagnosis of food allergies. Regardless of the causal mechanism the interview has the most important role in the diagnosis, and the gold standard is a double blind placebo controlled food challenge. Additional tests that can be performed in suspected IgE-mediated reactions include: skin prick tests, specific IgE measurement, component-resolved diagnostics and in doubtful cases basophil activation test (BAT). Due to the fact that the spectrum of the symptoms of the type I food hypersensitivity can include potentially life-threatening reactions, diagnosis is often limited to in vitro assays. In these cases BAT may play an important role--in a recent publication, for the first time BAT reactivity reflected the allergy severity and BAT sensitivity reflected the threshold of response to allergen in an oral food challenge. Atopy patch tests are valuable diagnostic tool in suspected type IV food hypersensitivity, but due to the lack of standardization they are not used routinely. The cytotoxic test has been developed on the basis of the observations that leucopenia developing in the type II hypersensitivity reaction mechanism may be one of the symptoms of food allergy. Unfortunately its use is not justified in any method fulfill the criteria of controlled clinical trial. Food allergy can also develop in the type III hypersensitivity reaction, but there is lack of research supporting the role of IgG measurement in the detection of allergens responsible for symptoms. Each result of additional diagnostic tests before the introduction of food elimination should be confirmed in double-blind, placebo-controlled or open food challenge, because non proper diet is not only ineffective but also can be harmful.

[Clinical symptoms of the allergic rhinitis and coexisting cross reactions in patients treated with the birch and/or grasses allergens against a background of the pollen exposure].

During specific immunotherapy (SIT) it is recommended to monitor the patient symptoms in relation to the exposure of allergen which the patient is treated. The aim of the study was to analyze the clinical symptoms of allergic rhinitis (AR) and cross reactivity in patients undergoing specific immunotherapy using pollen allergens (SIT) (birch and/or grasses) against the pollen exposure in 2014-2016. The study group, consisted of patients with pollen allergy, treated by SIT with birch and grasses allergens, who showed oral allergy symptoms. Patients of the control group were sensitive to birch and/or grasses allergens and they were not treated with SIT. The analyses were based on the results of questionnaires and patient symptom diaries. Timing of the pollen seasons of birch and grasses were similar, however the significantly higher concentration of birch pollen was found in 2014 and 2016, comparing to 2015. The strongest relationship between the pollen concentration and symptoms severity was revealed in the study group in patients desensitized by combined vaccine (birch/grasses), while in the control group, in patients sensitive to both taxa. In 35.3% of patients in the study group, a significant decrease in symptoms after foods was found, especially in patients desensitized with mixed vaccine (birch, grasses) after consumption of vegetables and in patients desensitized with grasses allergens after the fruits and nuts. SIT has a significant impact on the symptoms score reduction, which could be modified by the changeable seasonal pollen exposure.

Molecular methods of diagnosing allergic sensitization in patients with atopic dermatitis and infected by with Staphylococcus aureus.

The severity of allergic symptoms in patients with atopic dermatitis (AD) intensifies when the number of colonies patient's of Staphylococcus aureus on patents' skin increases. The basic feature determining the quality of any diagnostic test for S. aureus is its credibility. Performing a test always carries the risk of obtaining false positive and/or false negative results. Furthermore, producing material for microbiological analysis of internal body cavities is sometimes difficult. Therefore, in our study, we compared the results of three tests to determine if their results were mutually compatible and if they confirmed whether S. aureus was present in patients with AD and what was its role in the development of the disease in those patients. Infection with S. aureus was tested in patients with AD and healthy volunteers using the API Staph system. The specific IgE antibodies for staphylococcal enterotoxin A (SEA) and B (SEB) were measured using the UniCAP system. The secretion of IFN-γ, IL-2, IL-13 by peripheral blood mononuclear cells (PBMCs) after stimulation with SEA and SEB were studied with Elispot assay. We found that only certain patients with AD and S. aureus produced antibodies against SEA and SEB in the acute phase of AD. The secretion of IFN-γ was low in patients with exacerbated AD and S. aureus. Testing for the presence of S. aureus in the mucous membrane of the nasal vestibule and skin lesions is not sufficient for complex diagnosis of the role of S. aureus in the pathomechanism of AD. Measuring the presence of antibodies against bacterial components in patients' serum and the reactivity of patients' immune cells against these bacterial components is required in order to accurately diagnose this role of S. aureus in a patient.

Growing problem of allergy in population of children and adolescents living in Polish cities.

Allergy is a worldwide medical problem and prevalence of allergic diseases is on the rise. In the study a population of 7494 children and adolescents living in Cracow responded to a questionnaire based on the ISAAC study complemented with questions concerning food allergy. 49.1% of respondents reported symptoms suggestive of allergic disease. 52.7% of them (25.9% of respondents in total) had not received any treatment despite the reported symptoms. The most frequently reported allergic disease was allergic rhinitis followed by atopic dermatitis, asthma, food allergy and allergic contact dermatitis. In the adolescents group allergic rhinitis was followed by asthma. 487 voluntary applicants were further analysed in order to confirm specific allergic disease. Given the final diagnosis, allergic rhinitis was also the most common disease. Interestingly in both categories (preliminary and final diagnoses) we noticed that it is substantially more frequent in the older group which in our opinion can be linked to the impact of polluted urban environment but there is a need for further research in this area.

Drug-specific in vitro secretion of IFNγ in the diagnosis of drug-induced exanthemas: electrochemiluminescence assay versus previously used diagnostic methods.

The in vitro diagnosis of delayed drug hypersensitivity reactions remains a research problem. We measured drug-specific IFNγ release in peripheral blood mononuclear cells sampled from patients with drug-induced maculopapular exanthema and the age- and sex-matched control group. This is the first study to directly cross-compare an ultrasensitive assay based on an emerging electrochemiluminescence technology (ECL), the standard lymphocyte proliferation assay and three following tests detecting IFNγ at different steps of its production: intracellular in CD3+CD4+ cells (flow cytometry), secretion at the single cell level (enzyme-linked immunospot assay), bulk content in cell culture supernatant (enzyme-linked immunosorbent assay, ELISA). The highest rate of drug-positive responses were recorded for ELISA and ECL tests (56.25%). No false-positive responses were observed--all tests were negative in the control group. We demonstrated that IFNγ-detecting ELISA is not less efficient than ECL test, however, it is easily available and cheap, which makes it a potential method of choice in the future.

[Prevalence of drug allergy claims in school children]. / Czestosc zglaszania alergii na leki u dzieci szkolnych.

BACKGROUND: Adverse drug reactions, especially drug hypersensitivity reactions (DHR) are common problem, which impacts on prescription choices. In general pediatric populations data on the epidemiology of DHRs are scarce compared to in-patient and adult populations. In the current study, we assess the prevalence of parental-reported drug allergy in school children. METHODS: We performed a cross-sectional survey of a general 6- to 7-yr-old children population from Krakow. The life occurrence of drug allergy along with basic data on sensitization to common environmental allergens was assessed using a self-administered questionnaire. Four thousand eighty questionnaires were collected. RESULTS: The prevalence of drug allergy according to answers given by parents was 3.38% (138/4080). The most common reported drugs were penicillin and its derivatives (39%), followed by sulphonamides (25%), cephalosporins (9%) and non-steroidal anti-inflammatory drugs (8%). Sensitization to inhalant allergens, food allergens or haptens was not significant risk factor for drug allergy. CONCLUSIONS: The study showed that drug allergy claims in children are highly prevalent. Although results are based on parental perception, they point out how many patients need complete diagnostic work-up to confirm or exclude drug allergy. Otherwise they are treated with alternative drugs, which are usually less effective and/or more expensive.