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Birch (Betula pendula) pollen causes inhalant allergy in about 20% of human population in Europe, most of which is sensitive to the main birch allergen, Bet v1. The aim of the study was to find out (i) whether and how the analysed birch individuals differ in regard to composition of individual subunits of pollen proteins and to protein content in these subunits; (ii) whether the level of particulate matter relates to concentration of Bet v1 allergen. Study was performed in Southern Poland, in 2017–2019. Pollen material was collected at 20 sites, of highly or less polluted areas. Protein composition was analysed by SDS-PAGE, while the concentration of Bet v1 was evaluated by ELISA. The obtained results were estimated at the background of the particulate matter (PM10) level and the birch pollen seasons in Kraków. The electrophoregrams of pollen samples collected at different sites showed huge differences in staining intensities of individual protein subunits, also among important birch allergens: Bet v1, Bet v2, Bet v6 and Bet v7. The level of Bet v1 was significantly higher in the pollen samples collected at the more polluted sites. While the birch pollen allergenic potential is determined, the both pollen exposure and the content of the main allergenic components should be considered, as factors causing immunological response and clinical symptoms manifestation in sensitive individuals.

Introduction Hereditary angioedema (HAE) is a rare autosomal dominant disease caused by genetic dysfunction of C1 inhibitor (C1-INH) due to mutations in the SERPING1 gene. The disorder is mediated mainly by bradykinin. The clinical course of the disease is varied and not related to genetic changes. Objectives We aimed to evaluate redox homeostasis of peripheral blood mononuclear cells (PBMCs) in patients with HAE due to C1-INH deficiency (C1 INH HAE) by measuring the levels of reactive oxygen species (ROS) of PBMCs as well as plasma advanced glycation end products (AGEs) and advanced oxidation protein products (AOPPs). We also aimed to assess the effect of bradykinin on ROS levels. Patients and methods We enrolled 30 adults with C1-INH-HAE and 15 healthy individuals. The levels of ROS were measured by flow cytometry, while the plasma levels of AGEs and AOPPs were determined spectrophotometrically by enzyme‑ linked immunosorbent assays. Results Basal and hydrogen peroxide (H2O2)-induced ROS levels were higher in patients with HAE when compared with controls (P = 0.002 and P = 0.001, respectively), indicating abnormalities in redox homeostasis. Plasma AOPP and AGE levels were similar in both groups. Bradykinin reduced basal and H2O2-induced ROS generation in PBMCs only in patients with HAE (P = 0.03). Conclusions The higher basal and H2O2-induced ROS levels in patients with C1 INH HAE indicate redox imbalance. However, by reducing basal and H2O2-induced ROS levels, bradykinin shows antioxidant action in this disorder.

INTRODUCTION Hymenoptera insect stings (ISs) in the head and neck (HN) region are commonly considered to be a risk factor for grade IV systemic reactions (SRs) in patients with Hymenoptera venom allergy (HVA). However, clinical data addressing this issue are scarce. OBJECTIVES The aim of our study was to verify whether ISs in the HN region were related to a higher risk of grade IV SRs in patients with HVA. PATIENTS AND METHODS This retrospective cross‑sectional study included 195 patients aged 2 to 74 years and treated with venom immunotherapy due to at least a grade II SR to ISs. The study sample comprised 109 adults (56%; mean [SD] age, 41.08 [14.62] years; male, 50.5%) and 86 children (mean [SD] age, 9.53 [4.37] years; male, 72.1%; P0.001 for age and P = 0.002 for sex). The IS site was divided into 7 categories. RESULTS The HN region was the most common site for the IS (one‑third of the study group). In the entire study population, the risk of grade IV SRs was numerically, though insignificantly higher for ISs in the trunk (odds ratio [OR], 1.58; 95% CI, 0.42-5.92; P = 0.50) and legs (OR, 1.56; 95% CI, 0.49-5.10; P = 0.45), as compared with the HN region. The HN region showed a similar risk of grade IV SRs when compared with all the other IS sites combined into a single category (OR, 0.87; 95% CI, 0.43-1.75, P = 0.7). CONCLUSIONS ISs in the HN region were not confirmed to be a risk factor for grade IV SRs in patients with HVA, regardless of age and sex.

Background Aspirin desensitization (AD) is an effective and safe therapeutic option for patients with nonsteroidal anti-inflammatory drugs (NSAIDs)-exacerbated respiratory disease (N-ERD). The mechanisms driving its beneficial effects remain poorly understood. Objective To investigate the effect of long-term AD on clinical, biochemical and radiological changes in N-ERD patients. Methods The study group consisted of twenty-three individuals with N-ERD who underwent AD, followed by ingestion of 325 mg aspirin twice daily. Twenty patients completed the 52 weeks of AD. The following evaluations were conducted at baseline and in the 52nd week of AD: (i) clinical: asthma exacerbations, Asthma Control Test (ACT), Visual Analogue Scale (VAS) for the assessment of nasal symptoms; (ii) blood and induced sputum supernatant (ISS) periostin, (iii) phenotypes based on induced sputum (IS) cells, (iiii) ISS and nasal lavage (NL) concentration of prostaglandin D2 (PGD2), prostaglandin E2 (PGE2), tetranor-PGD-M, tetranor-PGE-M, 8-iso-PGE2, leukotriene B4 (LTB4), LTC4, LTD4 and LTE4, and urine LTE4. Results A significant improvement was observed in ACT (P = 0.02) and VAS score (P = 0.008) in the 52nd week of AD. ISS periostin and IS eosinophil count decreased significantly in the 52nd week of AD (P = 0.04 and P = 0.01, respectively). ISS and NL eicosanoid concentrations did not change following long-term AD. Conclusion and Clinical Relevance: AD is associated with a decrease in sputum periostin biosynthesis, which may prevent the recruitment of eosinophils into respiratory tissue and be one of explanation of the clinical benefits of AD. Long-term aspirin administration does not lead to an imbalance between pro- and anti-inflammatory ISS eicosanoids.

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.

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.

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.

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.

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.

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 (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.

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 was80 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 (humidity60%), 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.

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.

[b]Abstract Introduction and objective[/b]. Gluten proteins (gliadins and glutenins) are polymorphic wheat storage proteins of allergenic properties. Significant differences in chemical composition between both protein groups allow to expect highly specific immunological response of individual subunits and fractions in reactions with IgE sera of people allergic to wheat. The aim of these studies was to identify and characterize the most allergenic gluten proteins (GP) and nongluten proteins (NGP) occurred in two closely related wheat hybrid genotypes. [b]Materials and method.[/b] 3xC and 3xN wheat hybrids, which differ strongly in regard of gliadin composition, were analyzed. Seven people manifesting different symptoms of wheat allergy donated sera for the experiment. The technique of immunoblotting after SDS-PAGE was used for identification of allergenic subunits and fractions among GP and NGP. Immunologically active protein bands were visualized by chemiluminescence. [b]Results[/b]. Great variation of immunodetection spectra was observed. Results of immunoblotting showed LMW glutenins to be of highest, gliadins of medium, while NGP of lowest allergenicity for selected patients. The 43-kDa and 47-kDa LMW glutenin subunits, 40-kDa and 43-kDa γ-gliadin fractions and 49-kDa NGP can be considered as the most immunoreactive among all protein bands [b]separated by SDS-PAGE. Conclusion [/b] The observed differentiation of immunodetection spectra allows to model highly specific IgE-binding profiles of allergenic wheat proteins attributed to individual patients with symptoms of gluten intolerance. Highly immunoreactive subunits and fractions among GP and NGP were identified. The observed immunoreactivity of 49 kDa NGP is worth to emphasize, as it has never been reported as wheat allergenic protein before.

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.

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 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 kU A /L; interquartile range, 11.30-66.30 kU A /L) compared with adults (5.10 kU A /L; interquartile range, 2.03–8.30 kU A /L) ( P P 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.

Diagnosis and treatment of chronic urticaria (CU - Chronic Urticaria) is one of the most difficult issues in allergy practice. Studies on the etiopathogenesis of chronic urticaria suggest that environmental factors, immune, genetic and hormonal are involved in triggering and maintaining the reaction of hives. In practice, despite detailed diagnosis is usually not possible to determine the real cause of the disease. For this reason, the use of causal treatment for patients suffering from chronic urticaria is not possible and supportive treatment is often ineffective. 0.1% of the population suffers from chronic spontaneous urticaria, which main cause has not been detected. Very often CU coexists with autoimmune thyroid dysfunction. Studies confirm higher incidence of thyroid antibodies in patients with chronic urticaria than in the general population. We analyzed 100 patients who met the criteria for chronic urticaria according to the WHO, in Allergology Clinic of the University Hospital Jagiellonian University Medical College. According to our study 17.4% of patients with chronic spontaneous urticaria occurred elevated titer of anti-TPO antibodies. In patients with non-spontaneous type of CU, elevated titer of anti-TPO antibodies only occurred in 8.3% of patients, and this difference was statistically significant p =0.003. All the patients had thyroid levels normal and there were no clinical symptoms of thyroid dysfunction. Our research indicates a link with autoimmune thyroid diseases, especially Hashimoto's disease, with a prevalence of CU. It is necessary to conduct further studies to confirm the relationship of pathogenic autoimmune thyroid disorders with the occurrence of CU, which will help to provide the causal treatment of spontaneous CU coexisting with high levels of anti-TPO antibodies in patients with euthyroid status.

Recent remarkable findings of pharmacogenomics and basic scientific research provided insights in the pathogenesis of severe drug hypersensitivity reactions such as drug rush with eosinophilia and systemic symptoms (DRESS), abacavir hypersensitivity syndrome or blistering reactions (Stevens-Johnson syndrome, toxic epidermal necrolysis). There is increasing evidence for the strong associations of certain human leukocyte antigen (HLA) alleles with hypersensitivity to particular drugs. HLA genes may serve as genomic biomarkers of predisposition to severe adverse drug reactions and enable to prevent them. In this paper we review essentials and advances in this area.

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.

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.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.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.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.

Allergic diseases are important health problems. Medical history and results of additional tests play a crucial role in the diagnosis. Cytometric basophil activation test (BAT) is widely evaluated technique in inhaled, food, Hymenoptera venom and drugs allergy. The aim of our review is to systematize the available knowledge about possibilities of applying BAT, especially in the diagnosis of inhaled allergies. Basophil activation markers with proven sensitivity and specificity are CD63 and CD203c--each of them has its supporters and opponents among researchers. Use of the first one is more popular because of the direct relation of its appearance on the cell membrane of basophils with degranulation. The usefulness of BAT has been confirmed in the diagnosis of grass pollen, trees and house dust mites allergies in children and adults. BAT has a comparable to the routinely used tests sensitivity and higher (in most studies evaluated as 100%) specificity. The disadvantage of BAT that has been observed by some authors is "hyperreactivity" of basophils in atopic patients, leading to the appearance of false positives results. Basophil activation test seems to have a significant advantage over skin tests due to safety and lack of necessary to withdrawal antihistamines medications. Future research should focus on the use of BAT in diagnostically difficult cases and in monitoring of immunotherapy.

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.

Mastocytosis is a heterogeneous group of diseases characterized by excessive proliferation and accumulation of mast cells--in one or more organs. The number of symptoms and clinical prognosis vary depending on the disease. One of the most severe potential outcome of mastocytosis is anaphylactic shock. Early diagnosis and identification of triggers enables education and avoidance them. We describe the case of a 31-year-old woman with systemic mastocytosis (SM) without skin symptoms with multiple anaphylactic reactions, including two severe. Systemic mastocytosis was confirmed in bone marrow and genetic studies. We identify allergic triggers: latex and cefuroxime and also other non-immunological triggers as non-steroidal anti-inflammatory drugs (NSAIDs). The patient never had any allergic reaction after Hymenoptera stings. Only one result of serum tryptase was elevated. There was a need to determine the safety of antibiotic use and anesthetic drugs before cesarean. The moment when the disease was diagnosed and triggers were identifie, helped avoid further severe reactions.

Nasal mucosa cytology is an additional examination useful to differentiate chronic rhinitis in allergological and laryngological diagnostics, including chronic sinusitis. The aim of the study was to estimate the cytological picture in a group of patients with the highest percentage of squamous epithelia, suggesting the atrophic rhinitis. The analysis was carried out on the basis of cytological results performed in 3055 patients diagnosed because of chronic rhinitis. Among these patients, in 31 individuals the higher percentage (30% - 76%) of squamous cells was found. In six patients, the clear predominance of squamous cells over ciliated and goblet cells (3:1) was reported. Only in three patients the increased percentage in eosinophils was stressed (3%, 5% and 9% respectively). No basic and metaplastic cells were observed in the studied cytograms. The squamous cells occurred probably as a result of normal mucosa damages during the different inflammatory changes, both allergic and non-allergic, the impact of toxic agents, irritants (including nasal drops). The higher percentage of squamous cells in the nasal cytogram could have explained the low effectiveness of local inflammatory treatment in these patients.

According to asthma treatment guidelines the main goal of pulmonary rehabilitation is optimum asthma control. The Asthma Control Test (ACT) is a standardised five-item questionnaire for the assessment of asthma control. The study compares the pre- and post-treatment (subterraneotherapy) ACT score with other conventional respiratory parameters. The study included 21 patients with bronchial asthma who underwent a 3-week long subterraneotherapy programme in the 'Wieliczka' Salt Mine. The patients completed the ACT questionnaire before and 2 weeks after subterraneotherapy. At the same time, they underwent testing for concentration of nitric oxide in exhaled breath (FENO), peak expiratory flow (PEF) and spirometry. Subterraneotherapy influenced significantly the change of MEF75 (p = 0.03255) and improvement of ACT score (N = 21, p = 0.0016). The differences in other parameters were not statistically important, but in the case of FEV1 and VC parameters, the differences found were close to the statistical significance (0.05p0.10). The differences of results before and after subterraneotherapy were higher in the group assigned as patient with poor or moderate asthma control (ACT score20, N = 10) than in the group with good control of asthma (ACT score ≥ 20, N = 11). For parameters FEV1, VC, MEF75 and ACT score the improvement was significantly higher in the poor control group. The pulmonary rehabilitation programme combined with subterraneotherapy helps to control asthma especially in patients suffering from poorly or moderate controlled asthma.

UNLABELLED INTRODUCTION Cytotoxic mechanisms are present in the majority of delayed drug hypersensitivity reactions, but are not used as a diagnostic tool. OBJECTIVES The aim of the study was to compare cytotoxic‑based assays with a proliferation assay and drug patch tests in patients with maculopapular eruptions induced by antiepileptic drugs. PATIENTS AND METHODS Peripheral blood mononuclear cells of 23 patients and 24 controls exposed to the drugs were cultured under defined conditions. A drug‑specific response was assessed by measuring granzyme B (GrB) release with an enzyme‑linked immunospot assay, intracellular expression of granulysin (Grl) in CD3-NKp46+ cells with flow cytometry, perforin concentrations in cell culture supernatants with an enzyme‑linked immunosorbent assay, and using the lymphocyte proliferation test. Patch tests with culprit drugs were done in all patients. RESULTS Lymphocyte proliferation, GrB release, and Grl expression were significantly higher in patients than in controls, while perforin concentrations were not elevated. The sensitivities were 30.4%, 55%, 39.1%, and 17.4% for proliferation, GrB, Grl, and perforin‑based assays, respectively. A significantly higher rate of positive results was observed when assays were done within 2 years after a drug‑induced reaction. The specificities of all assays remained in the range of 95.8% to 100%. The results of patch tests were positive only in 3 patients (sensitivity, 14.3%) and negative in all controls. CONCLUSIONS In vitro assays based on the detection of Grl, and in particular of GrB, are superior to routine diagnostic tests in patients with hypersensitivity to antiepileptic drugs. They can detect a low‑level response that might be overlooked by standard techniques. In the remission phase, drug‑specific cells are more easily detectable directly in the circulation than in the skin.

[b]introduction and objective[/b]. Numerous epidemiologic studies have reported increased risk of allergic rhinitis and asthma in relation to ‘western life-style’, which represents diversity of factors. We hypothesized that residential proximity to major roadways, reflecting an exposure to traffic-related air pollution, is associated with prevalence of allergic respiratory symptoms in children. [b]materials and methods[/b]. A total of 8290 individuals of two age groups: 16 year olds and 7 year olds from Krakow, Poland were included. We used the Polish version of the International Study of Asthma and Allergy in Childhood supplemented with a question concerning the distance between a responder’s house and a high traffic density road: below 200 m, from 200–500 m, or more than 500 m. [b]results[/b]. Children and adolescents with a residential proximity closer to a major roadway had more frequent asthma-related symptoms in the last 12 months and at any time in the past. Consistent with the increased frequency of asthmatic symptoms, responders residing within 200 meters complained more often of sneezing, runny or blocked nose accompanied by itchy-watery eyes and hay fever in comparison to responders who resided 200–500 meters from a major roadway. The lowest rate of nasal symptoms was observed in residents living in the distance to major roads (> 500 meters). The rate of positive answers decreased in a distant-dependent manner. [b]conclusions[/b]. Our findings suggest an important spatial relationship between the distance from a major roadway and the evaluated respiratory symptoms. The results emphasize the need for more comprehensive air quality policies within urban areas with increased motor vehicle density.

Adverse drug reactions in children are serious public health problem. The overall incidence of ADRs is estimated at about 9.5% in hospitalized children and about 1.5% for outpatient children. ADRs have very diverse manifestations from minor skin rashes to potentially life-threatening severe skin reactions such as: acute generalized exanthematous pustulosis, Stevens-Johnson syndrome, toxic epidermal necrolysis and anaphylactic reactions. The aim of the study was to analyze the results of ADRs monitoring with particular regard to life-threatening reactions in the pediatric population in Poland. We analyzed spontaneous reports of ADRs in Poland in patients aged 0-18 years in the period 01.01.2004-31.03.2013. Each reported case was evaluated and classified by a clinical pharmacologist and an allergologist. Analysis of the data showed the occurrence of ADR in 846 pediatric patients including one case of Stevens-Johnson syndrome, and 90 (10.6% ADR) drug reactions classified as anaphylactic reactions (44 responses (5.2% ADR)) and non-immune anaphylaxis (46 responses (5.4% ADR)). Classes of drugs most frequently causing anaphylactic reactions were: anti-inflammatory and antipyretic (31%), antibiotics (19%) and drugs used in anesthesia (16%). The substances most commonly causing anaphylactic reactions/non-immune anaphylaxis were: paracetamol and lidocaine (with 8 responses). The most common clinical manifestations of ADRs are skin reactions (402 responses, 47.5%). Acute drug reactions in the pediatric population may have life-threatening clinical manifestations. Drugs that most commonly cause anaphylaxis are: anti-inflammatory/ antipyretic drugs (acetaminophen, aspirin, ibuprofen), antibiotics (cephalosporins, amoxicillin, sulfonamides), local anesthetics (lidocaine). Early diagnosis of allergy and choice of safe product can prevent life-threatening drug reactions.

Specific allergen immunotherapy should be modified according to sensitivity of the patient and the time interval between injections and seasonal allergen exposition. The aim of the study was to check the effectiveness of the multinomial logistic regression models predicting the pollen concentration during the pollen season in the immunotherapy trial in patients treated with grass and birch allergens. The study was performed in Krakow in 2011-2013. Models were validated for 2012 and 2013. The effectiveness of the total correct predictions slightly differed depending on the time series, in case of birch pollen the similar percentage of correct predictions was found in both study year, while in case of grass pollen, the predictions were more correct in 2012. A group of patients treated with grass and birch allergens filled in the diary cards during the pollen season. After the 2011 season 14 diary cards were analysed, while 18 and 19, in 2012 and 2013, respectively. Because of manifested symptoms, the injection dose was reduced during the season in 12 patients in 2011, in 9 patients in 2012 and in 6 patients in 2013. No visits were delayed because of medical indications. In some cases patients got the injection in time of the high pollen occurrence (2 cases, in 2011 and 2012). In 2013 in 10/17 patients the high pollen exposure was avoided thanks the information from pollen monitoring, in opposite to 1 and 8 patients in 2011 and 2012, respectively. Patients used antihistaminic drugs on request. The regional pollen monitoring data and satisfied co-operation with patients makes the possibility of closer control of the injection doses administration during immunotherapy in the pollen season.

According to scientific societies guidelines the indication for venom immunotherapy is based on the clinical history of the patient. Diagnostic tests, like skin prick test or specific IgE serum estimation are conduct to prove IgE dependent mechanism of allergy and insect identification. Recent guidelines indicates for group of patients with severe systemic reactions as a candidates for diagnostic testing and in consequence for immunotherapy. In some countries diagnostic tests are also performed in patient who have a history of large local reactions, if they are considered as a candidates for immunotherapy. Double sensitization in cases of patients unable to identify the culprit insect is a diagnostic and therapeutic problem. In our group of patients (n = 113) we confirmed the double sensitization in 30% cases. The addition of a major allergen labeling reduced the number of people actually double-sensitized to 8.84%. It was observed that in patients who are not able to identify insect double sensitization phenomenon is particularly frequent as much as 45.5% and in the determination of the major allergens in 18.2%. Such patients needed detailed diagnosis and in many cases the use of two vaccines to conduct immunotherapy.

Gliadins and glutenins--the main components of wheat gluten--are highly complex and polymorphic proteins of wheat kernels. They are also allergenic proteins causing a range of food allergies. We hypothesized that the diversity of chemical structures and properties may relate to the diversification of immunoreactive properties of various subunits and fractions of gluten proteins. In the present study we used IgE sera, obtained from patients manifesting different symptoms of wheat allergies, for immunobloting analysis, to prove the specificity of immunological reaction between IgE antibodies and individual gliadins and glutenins, separated by SDS-PAGE. The results suggest that patients have different characteristics of IgE binding to the separated protein subunits and fractions. Sera of two patients showed strong binding of omega-gliadins, while alpha-gliadins and LMW glutenin subunits of MW = 43 and 45 kDa were highly allergenic for two other subjects in the test group of patients.

Drug hypersensitivity can occur with almost any drug and may range widely in clinical severity from mild pruritus, acute urticaria or angioedema to life-threatening anaphylaxis. Affected patients usually avoid the suspected drug in the future, but in selected cases the particular drug is essential for optimal therapy due to unavailable or ineffective alternative therapy. Under these circumstances, desensitization may be performed. Desensitization protocols have been developed and are used for antibiotics, sulfonamides, non-steroidal antiinflammatory drugs, insulins, biologic agents, and many others. Desensitization procedure is based on the induction of a temporary state of tolerance of a substance responsible for a hypersensitivity reaction. Gradually increasing doses of the drug are administered to the patient over several hours to a few days, until the total cumulative therapeutic dose is achieved and tolerated. Hypersensitivity to acetylsalicylic acid is a common indication to desensitization in a daily practice. A few protocols for this drug have been described. Recently, 7 patients hypersensitive to acetylsalicylic acid have been desensitized in our department due to cardiologic and rheumatologic reasons. In this group, desensitization procedures were performed successfully and the patients could continue pharmacotherapy with aspirin.

Nasal exfoliative cytology is a complementary tool in diagnostics of allergic (AR) and non-allergic (NAR) rhinitis. The aim of the study was to determine the usefulness of the nasal cytology in patients sensitive to common inhalant allergens with positive SPT(+) and negative SPT(-) (Skin Prick Tests) depending on the symptoms of intermittent and persistent rhinitis. The study was performed in a group of 285 patients treated in the Department of Allergology University Hospital in Krakow, suspected on AR in 2008-2010. The patients were made a smear test of inferior nasal concha. The samples were stained using the eosin-hematoxylin method and examined under a light microscope (1000x). Patients were divided into two groups: SPT(+) (144 patients) and SPT(-) (141 patients). Depending on the percentage of obtained eosinophils each group was divided into three subgroups: 0-2%, 3-20%,20%. In the most percentage of patients with 3-20% of eosinophils in nasal cytology were found, in both studied groups (SPT)(+) and (SPT)(-), while the highest percentage of eosinophils (20%) was observed in the bigger group of patients with SPT(+), than with SPT(-). The number of patients with eosinophils20% in the SPT(+) group was higher in patients with persistent symptoms (NS differences), while in the SPT(-) group, the number of patients with intermittent symptoms in the subgroup20% of eosinophils statistically prevailed (p0.001). The mean percentage of eosinophils in both groups was comparable, while the statistically significant differences were found considering the distinguished subgroups. In intermittent SPT(+) group the most sensitizing allergens were pollen grains (birch or grass pollen), while the patients with persistent AR symptoms were mainly sensitive the house dust mites. The mean percentage of eosinophils in an exfoliative cytology correlated significantly with allergic rhinitis symptoms and SPT results, the most evident relationship was found between higher level of eosinophils and the patients with confirmed AR diagnosis on the basis of positive SPT, manifesting the intermittent symptoms.

The symptoms of pollen allergy in the European population occur in a period of increased pollen precipitation, and take the form of allergic rhinitis and conjunctivitis, bronchial asthma, contact urticaria, and food allergy. Diagnosis in addition to medical history, takes into account the positive results of skin tests and elevated allergen-specific IgE antibodies (specific IgE) in serum. These studies are considered to be objective diagnostic tests confirming the diagnosis of pollen allergy. Not in every case there is a correspondence of symptoms and results of diagnostic tests, which puts into question the accuracy of the diagnosis of pollen allergy. The aim of this study was to test the characteristics of patients with oral allergy syndrome on the background of all patients with pollen allergy and evaluation of the diagnostic value of history, skin tests and specific IgE levels in the diagnosis of patients with pollen allergy and oral allergy syndrome. A retrospective analysis of the cases of 85 patients with a diagnosis of pollen allergy and the 30 patients with OAS was performed. In our study the most common sensitizing allergen in patients with OAS was birch pollen, while patients showing no symptoms of OAS were equally sensitive to timothy and birch pollen. The main food responsible for the presence of the OAS in the mechanism of cross-allergy to pollen was an apple. Among patients with OAS we did not show significantly higher incidence of polyvalent allergies. It was shown, however, that there is a tendency that the maximum concentration of allergen-specific IgE causing clinically significant symptoms, ie in line with the pollen season, is higher in the OAS patients than in the absence of OAS. Further research is needed using new diagnostic methods, which would predict future symptoms after eating certain foods in particularly endangered patients with pollen allergy.

Hymenoptera venom allergy is related to higher risk of potential life -threatening anaphylactic reactions, which leads to anxiety and decreased quality of life.The aim of this paper was: 1) estimation of fear level of being re-stung among venom allergy adults treated with venom specific immunotherapy (VIT)--before and during treatment; 2) estimation of expectation of outcome of VIT as compared to level of anxiety of being re-strug, in the Visual Analogue Scale--VAS score; 3) identification of factors influencing changes in the fear level among patients during VIT.The study group comprised 42 patients (18 women, 24 men) in the mean age 42.6 years, with bee or vespid allergy, who had been qualified to the VIT treatment with Alutard SQ. Visual Analogue Scale--VAS and the Expectation of Outcome Questionnaire were used. The demographic data were collected.The VAS score before VIT for insect venom allergic patients was 8.8 (SD = 0.9). It decreased after achieving maintenance dose to 3.1 (SD = 1.6) and was significantly lower in men (p0.05). Score achieved in the Expectation of Outcome Questionnaire was for each question 2.2 (SD = 1.5) and there was correlation with VAS score during VIT.The patients with insect venom allergy, who undergo a serious allergic reaction (SR) as a result of being stung and who are qualified to VIT, have a high level of anxiety of being re-stung. Achieving the maintenance dose of VIT, results in a significant decrease of anxiety level in women and men, significantly so in men. There is a significant correlation between VAS score and the Expectation of Outcome Questionnaire results during VIT. Both VAS for anxiety level and the Expectation of Outcome Questionnaire can be simple, easily available and useful instruments helping to estimate quality of life. VIT significantly decreases the patients level of anxiety of being restung and improves their quality of life.

Routine qualification for specific immunotherapy (SIT) is based on clinical history and skin prick tests (SPT) or specific IgE (sIgE). In cases of discordance between these two, basophil activation test (BAT) may be decisive. The aim of the present study was to determine the specificity and sensitivity of BAT, sIgE, and SPT, and to analyse cases, in which clinical data and SPT alone would result in wrongful qualification for SIT.BAT results and sIgE levels to Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df) were determined in 52 pediatric patients qualified for SIT based on clinical history and positive SPT. The group included 21 children qualified for SIT with birch or timothy grass, used as reference for specificity and sensitivity calculations for BAT, sIgE and SPT.The sensitivity and specificity of BAT, using SPT as "gold standard" was 96.9% and 88.9% for Dp, and 89.3% and 100% for Df, respectively and the sensitivity and specificity of sIgE were 89.7%, 88.9% for Dp, and 92.9% and 94.4% for Df. When using BAT as "gold standard", the sensitivity and specificity of SPT was 90% and 90.5% for Dp, 92% and 84,6% for Df, and these indices for sIgE were 87.1% and 90.5% for Dp, 100% and 87.5% for Df. BAT did not confirm the initial qualification for SIT in 2 patients, revealing an unspecific basophil activation. Negative nasal provocation test ultimately confirmed the false-positive SPT, which could be explained by the co-existence of urticaria in those children. In further 2 children qualified for SIT with timothy and birch, BAT revealed lack of reactivity to respective allergens. Altogether BAT helped in avoiding unnecessary SIT in 4 out of 52 children (7.7%).In most cases, SPT, sIgE and BAT provide comparable information, however, SPT results care is advised in patients with co-existing urticaria. BAT is useful in verifying the actual relevance of allergens selected for SIT and helps in avoiding long-lasting, arduous, costly, and ineffective immunotherapy of wrongly qualified cases.

Contact allergy is detected in every second child with the symptoms of chronic or recurrent eczema, and in every third child the final diagnosis is allergic contact dermatitis. Haptens responsible for the majority of contact sensitizations in children are substances ubiquitous in our environment, e.g. metals, preservatives, fragrances, propolis, and balsam of Peru. Much concern is provoked by the higher rates of sensitization to fragrances in younger children, compared to adolescents, which may be attributed to the higher exposure nowadays of infants and children to fragrant products. On the other hand, a limitation of exposure to the preservatives thimerosal and Kathon CG has resulted in decreased rates of sensitization to these haptens. Altogether, these observations demonstrate that the rates of contact sensitizations in children reflect changes in their environment, and limitations imposed on the use of haptens with strong sensitizing properties, may be an effective tool in the prevention of contact allergy.

Systemic sclerosis (SSc) is complicated by pulmonary hypertension and right ventricle (RV) failure in approximately 10% of the patients. Factors influencing the reactivity of pulmonary circulation to vasodilators are not established, while the examination of vasoreactivity is important in determining the treatment, because systemic administration of oral vasodilators can induce severe adverse events in nonresponders. The mechanism of RV failure in SSc is unclear and may result either from increased RV afterload or intrinsic myocardial disease. The aim of the study was to assess the reactivity of pulmonary circulation to inhaled nitric oxide (iNO) and to evaluate its influence on RV function in SSc patients with elevated right ventricle systolic pressure (RVSP). In 60 SSc patients aged 24–73 (58 females, two males; 33 patients with limited SSc and 27 with diffuse SSc), echocardiographic examination with tissue Doppler echocardiography (TDE) was performed. RV function was measured by systolic (S) and early diastolic (E) velocity of tricuspid annulus by TDE. In patients with RVSP >45 mmHg, the reactivity of pulmonary circulation was assessed by iNO test. High-resolution computerized tomography (HRCT) was performed to assess the extent of pulmonary fibrosis. Of 14 SSc subjects with elevated RVSP (13 females, one male; RVSP 47–62 mmHg), positive reaction to iNO was observed in five (RVSP decreased from 51.6 ± 3.7 to 32.24 ± 2.3 mmHg); nine patients were not reactive (RVSP 53.5 ± 5.7 mmHg before iNO vs. 49.6 ± 6.7 mmHg). RV systolic function was decreased in patients with elevated RVSP as compared to the patients with normal pulmonary pressure (S velocity 13.2 ± 1.3 vs. 14.4 ± 1.6 cm/s, respectively, p

The differential diagnostic work-up of children with chronic eczema should involve patch testing, also in cases with confirmed atopy. In our previous study, contact allergy was detected in every second child with chronic eczema. The aim of the present study was to identify the most important sensitizers in atopic children with eczema. During an allergy screening program, 103 consecutive children aged 7-8 and 93 adolescents aged 16-17 were enrolled. The inclusion criterion was chronic recurrent eczema as detected with the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire and atopy, defined as positive skin prick test to one or more common airborne or food allergens. The children were patch-tested with the newly extended European Baseline Series (EBS, 28 test substances) supplemented with propolis, thimerosal, benzalkonium chloride, and 2-phenoxyethanol. In total, 67.0% children and 58.1% adolescents were found patch test positive. Among children, 35.9% reacted to nickel, 16.5% propolis, 11.7% thimerosal, 9.7% cobalt, each 6.8% fragrance mix (FM) I and chromium, and 5.8% to FM II. Among adolescents, 37.6% reacted to thimerosal, 19.4% to nickel, 6.5% to cobalt, and 5.4% to propolis. We demonstrate the advantage of using FM II - a new addition to the EBS that detects a relatively high proportion of contact hypersensitivity among children. An important sensitizer from outside EBS is propolis, which according to the frequency of sensitization occupies rank 2 in children and rank 4 in adolescents. These data show that propolis should be included into routine patch testing in children.

Contact allergy is the most frequent type of allergy, affecting 26-40% of all adults and 21-36% children. The gold standard in the diagnosis of contact allergy is patch test.To study the influence of the range and composition of patch test series on the efficacy of the diagnostic procedure.Retrospective analysis of the frequency of positive reactions among patients diagnosed with patch tests at our Department during 2 periods: From December 2003 to March 2005, patients were tested with a series of 9 substances plus white petrolatum as the negative control. From April 2005 to July 2008, the series was expanded to 21 substances, while petrolatum was removed.In the analyzed period, 1379 patients were tested with 9 substances plus petrolatum (group referred to as "G9") and 682 patients with 21 substances ("G21"). In G9, at least one positive reaction was observed in 343 (24.9%, 95% CI: 22.6-27.2%) patients, as compared to 376 (55.1%; 95% CI: 51.4-58.7%) in G21 (p0.0001). The increase in the number of tested substances from 9 to 21 led to significant increase in the mean number of positive reactions per one patient (0.34 in G9 versus 0.90 in G21; p0.0001). We have not observed any positive reaction to white petrolatum.Patch testing with more extensive test series increases the chance for the detection of patient's sensitizations. As we have not observed any positive reaction to white petrolatum, using the vehicle as negative control does not seem to offer any advantage.

Traditionally, allergic contact dermatitis (ACD) has been associated with the activity of Th1 lymphocytes that secrete interferon gamma. Recent evidence indicates that other cells, e.g. interleukin 5 (IL-5)-secreting Th2 or Tc2 cells may be among the key effectors of ACD. The aim of the present study was to assess the influence of nickel-specific IFN-gamma secretion (marker of Th1 and Tc1 activity) and IL-5 secretion (Th2 and Tc2) on the clinical outcome (patch test score) in nickel-allergic patients. 40 women with suspicion of ACD were involved, aged from 14-54 (median 31.5) years. They were patch tested with NiSO(4). Peripheral blood mononuclear cells (PBMC) from the patients were cultured and analysed for IFN-gamma and IL-5 secretion in response to NiSO(4). A series of statistical models (classical logit or cloglog link function) were used. We demonstrate that nickel-specific IL-5 secretion by PBMC is correlated with the intensity of patch test reaction (p=0.05), with no significant effect of IFN-gamma. An increase in the nickel-specific IL-5 secretion from PBMC by 10 pg/ml is associated with a 10-20% increase (depending on statistical model) in the odds ratio of the patient to have a higher patch test score. These findings support the assumption that cells secreting IL-5 (e.g. Th2, Tc2) play a more important role in the pathogenesis of ACD than previously thought.

Background: Patch testing is an essential procedure in the investigation of eczema in children. Objectives: To analyse the frequency of contact hypersensitivity and allergic contact dermatitis among Polish children with eczema. Patients/methods: During an allergy screening programme involving 9320 children aged 7 and 16 years, 12.6% reported symptoms of chronic/recurrent eczema. From this group, a representative sample of 229 eczema children underwent patch testing: 96 children aged 7 years and 133 teenagers aged 16 years. Patch testing was with 10 allergens: methylchloroisothiazolinone/methylisothiazolinone (MCI/MI), nickel sulfate, mercury ammonium chloride, thimerosal, cobalt chloride, potassium dichromate, lanolin, fragrance mix I, Myroxylon pereirae (balsam of Peru), and colophonium. Results: 49.4% tested children were found patch test (PT) positive. 43.8% of 7 year olds with eczema were PT positive, with sensitization to nickel sulfate (30.2%), thimerosal (10.4%), cobalt chloride (8.3%), fragrance mix I (7.3%), MCI/MI (6.3%), potassium dichromate (6.3%), M. pereirae (3.1%), mercury ammonium chloride (2.3%), and colophonium (1.0%). 52.6% teenagers were PT positive, with sensitization to nickel sulfate (23.3%), thimerosal (27.8%), cobalt chloride (10.5%), potassium dichromate (6.0%), mercury ammonium chloride (2.3%), M. pereirae (1.5%), and MCI/MI (0.8%). The final diagnosis of allergic contact dermatitis was confirmed in 36% of 7 year olds and 26% of 16 year olds. Conclusions: Every second child with eczema is PT positive, whereas every third child is finally diagnosed with allergic contact dermatitis.

It is very difficult to differentiate Melkersson-Rosenthal syndrom and macrochelitis granulomatosa. Two neurologists: Ernt Melkersson and Curt Rosenthal in 1928 and 1931 described classical symptoms of this syndrom: recurring facial paralysis, swelling of the face and lips and the development of folds and furrows in the tongue. In 1945 Miescher described a few cases of primary interstitial inflammation of the lips and gave them the name macrochelitis granulomatosa. Some patients had paralysis of the facial nerve, others folds of the tongue. In the present study the etiopatho-genesis, diagnostic and therapeutic problems are presented.

Type IV of hypersensitivity reaction is usually manifested in the skin in different clinical pattern. According to traditional Gell and Coombs classification, the mechanism of IV type of allergic reaction has been associated with contact allergy with the activity of lymphocytes Th1 secreting interferon gamma. Now, this vision seems to be too simplified. In the last years there were publications, which can throw a new light on these complicated mechanisms leading to the development of the type IV of allergy, especially to drugs, nickel and other haptens and also can explain the differentiation of clinical pattern in respective patients. The skin symptoms in type IV of hypersensitivity are triggered by activation of specific T-cell CD4+ and CD8+. Immunohistochemical and functional analysis of reactive T-cell has shown that the delayed hypersensitivity reaction depends on the secreted cytokines. For example maculo-papular exanthema may be either triggered by Th1 or Th2 in nature and cytokines interferon gamma, tumor necrosis factor alfa or interleukin-4, 5 and 13. Bullous reactions (i.e. Stevens-Johnsons Syndrome or toxic epidermal necrolysis) are characterized by widespread keratinocyte apoptosis, a consequence of high CD8+ T-cell involvement and the molecular cytotoxicity of Fas, perforin and granzyme B. Pustular exanthema reactions are stimulated via the T-cell release of 11-8 and granulocyte-monocyte colony-stimulatig factor (GM-CSF). For the better understanding of these inflammatory cascades deleted type IV of hypersensitivity reactions have been re-classified into four main subtypes: 1. IVa with Th1 and monocyte directed and cytokines: IFNgamma, IL-1, IL-2, 2. IVb with Th2 and eosinophils directed and cytokines: L-5, IL-4, IL-13, 3. IVc with T CD8+ directed and cytokines: perforin, granzyme B, Fas Ligand, 4. IVd with T CD4+, CD8+ and neutrophil directed and cytokines: IL8, GM-CSF. Clinically delayed hypersensitivity eruptions are often an overlap of cytokine pathways, with one preferential reaction dominating the final picture. Type IVa and IVc play a role inthe mechanism of contact dermatitis, however type IV b in chronic asthma, chronic allergic rhinitis and maculo-papular exanthema with eosinophilia, type IV c in bullous reactions (i.e. Stevens-Johnsons Syndrome or toxic epidermal necrolysis), so type IV d in pustular exanthema reactions (i.g. AGEP - Acute Generalized Exanthematosus Pustule, Behcet disease). This different clinical pattern of allergic disease mainly including drug allergy to nickel and other haptens as well as chronic asthma and allergic rhinitis may be explained by above mechanisms. The study of different mechanisms of four subtypes of type IVof allergic reaction may be helpful in the differential diagnostics and in the treatment of allergic diseases.

Nickel allergy constitutes a serious health problem of modern societies. Hypersensitivity to this metal is found in 13% adults and 8% children. Risk factors for nickel allergy are: female gender and early exposure to nickel, e.g. piercing. Various mechanisms of inducing nickel allergy are possible, which is also reflected in the different clinical pictures. Nickel can induce allergic reaction in 3 different ways: 1) it binds to carrier protein in the extracellular space and subsequently is processed and presented by antigen presenting cell (APC) in the context of MHC class II molecule, which activates CD4+ lymphocytes, 2) Ni penetrates into the cell where it binds to intracellular proteins, and subsequently it is presented in the context of MHC class I molecule, which activates CD8+ lymphocytes, 3) Ni can "bridge" MHC molecule together with the TCR receptor on lymphocyte without actually filling the antigen-binding site, which is in analogy to superantigens. Both Th2/Tc2 (IL-4, IL-5, IL-13) and Th1/ Tc1 (IFNgamma) take their part in the development of contact allergy to nickel. The trafficking of the effector cells to target organs (where the inflammatory reaction actually takes place) is controlled by homing antigens and chemokine receptors that are expressed on their surface. The accumulation of effector cells in a target organ can determine the symptoms of nickel allergy (the skin, mucosa etc.). The acquisition of nickel tolerance is possibly dependent on the IL-10 secretion by specific lymphocytes.

Nickel is knows as the most common cause of allergic contact dermatitis, as well as diffuse eczema, allergic rhinitis and bronchial asthma. The mechanism of contact allergy to nickel is well known. In spite of numerous investigations, the mechanism of systemic allergy to nickel is still not clear. 22 patients with positive patch tests to nickel were analyzed. On basis of clinical symptoms the patients were divided into two groups: 1. with contact allergy dermatitis to nickel--8 patients 2. with systemic allergy to nickel (allergic rhinitis and/or diffuse eczema--14 patients. The control group included non-atopic patients with negative patch test to nickel--6 patients. 10 ml of blood were taken from each patient and peripheral mononuclear blood cells (PMBC) were isolated. In PBMC culture, NiSO4 and PHA were stimulated. The control group was non-stimulated cells. The supernatants were collected after 3 and 6 days of culture and the levels of cytokines IL-5, 4 and IFNgamma were measured (ELISA). The concentration of IFNgamma in supernatants from stimulated as well as non-stimulated cells from patients with contact allergy to nickel was higher in comparison to the control group. The concentration of IL-5 in this group was low. There was an increase in the production of IFNgamma and IL-5 after NiSO4 stimulation in patients with systemic allergy to nickel. The higher concentration of IFNgamma in the same groups of patients investigated was in supernatants from the third day of PBMC culture were compared to the sixth day. After 3 and 6 days of culture, the concentration of IL-4 (ELISA) was below detection level in all supernatants analyzed.IFNgamma plays an essential role in the mechanism of developing of contact allergy to nickel; and IFNgamma as well as IL-5 play a role in the mechanism of developing systemic allergy to nickel. The third day of PBMC culture is more reliable for IFNgamma estimation.

Eosinophiles are the cells, which play a crucial role in different pathological events. The atopic and parasitic disorders, as well as hipereosinophilic syndrome (HES) are the most typical processes in which they take part. Eosinophilia, which is the rise of number of eosinophiles in peripheral blood above 500 in 1 microl, or above 5% in smear of leucocytes can exist both in allergic and infectious diseases, as well as in neoplasmic and autoimmunological disorders, and in reactions to drugs or others. The mechanism of activation of eosinophiles in atopic diseases is well known and is connected with IL-5, IL-4 and IL-13. According to the last literature data, three routes of activation of eosinophil can be distinguished: Th2 route connected with cytokines: IL-5, IL-4 and IL-13, Th1 route connected with cytokines: IFN-gamma 11-12, GM-CSF, as well as internal route connected with IL-1, TNF, GM-CSF. The experience of different routes of eosinophil's activation may be helpful in the differential diagnosis of eosinophilia.

Eosinophilia in peripheral blood occurs in allergic as well as non-allergic diseases. In the research there were 26 patients with eosinophil level above 5% encountered in leukocyte smear of peripheral blood encountered. Patients were split into 3 groups: patients with allergic atopic disease (ANN), patients with non-atopic (contact dermatitis) disease and patients with upper airways infection and/or parasitic disease. In the researched groups there were no statistically significant differences of ECP level in serum and of ECP/Eo ratio detected. There were also correlations between ECP and ECP/Eo ratio for every researched group analyzed. The highest correlation was observed in patients with contact dermatitis and in non-atopic patients in general. Initial results show uselessness of applied research methods in the routine differential diagnosis of eosinophilia.

Prevalence of allergic diseases has been increasing over recent decades, especially in developed countries. The aim of this study was to assess prevalence of bronchial asthma and allergic rhinitis in children of Krakow at school age, depending on home and school environment conditions. The study was carried out in two stages: questionnaire based on ISAAC questionnaire and skin prick tests in children with symptoms suggesting allergic disease were performed. Altogether 532 children from 4 primary schools (2 in central and 2 in peripheral parts of the city) were included. Symptoms suggestive of allergic disease were reported by 304 children (57.1%), including asthma in 145 (27.2%) and allergic rhinitis in 288 children (54.1%). Asthma and allergic rhinitis was previously diagnosed in 19 (4%) and 138 (26%) children respectively. In younger age groups more children reported symptoms suggestive of asthma. Pupils from schools in the center of Krakow exposed to high level of traffic significantly more frequently reported asthma and had diagnosed allergic rhinitis. Increased prevalence of allergic rhinitis symptoms was observed in children exposed to passive smoking. Skin prick tests were positive only in 29.7% of children qualified for them. The results of the study suggest that in pupils with allergic rhinitis in whom skin prick tests are weakly positive for inhaled allergens, environmental factors may play an important role in the development of clinical symptoms.