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.

Clinical Characteristics and Quality of Life in a Cohort of Polish Pediatric Patients with Hereditary Angioedema.

Hereditary angioedema (HAE) is a rare genetic disease. It is characterized by recurrent attacks of angioedema. Evidence to what extent it affects patient functioning is limited in the pediatric population. We aimed to determine the clinical characteristics and management of Polish children with HAE and to measure the health-related quality of life (HRQoL) of these patients. This cross-sectional study was conducted among 21 pediatric patients and their caregivers, as well as 21 respective controls randomly selected from the general population. During routine follow-up visits, standardized pediatric quality of life questionnaires (PedsQLTM 4.0) were administered to all caregivers and adolescents (≥13 years). Caregivers also completed a structured medical interview regarding the clinical characteristics and treatment of children with HAE during the previous six months. During this period, 57% of patients had low (group I), 24% moderate (group II), and 19% high (group III) HAE activity, corresponding to ≥10 attacks per 6 months. None of the patients received long-term prophylaxis. The children in group III had a lower HRQoL than other groups and controls on all dimensions of the PedsQLTM 4.0. The lowest scores in all groups were observed in the emotional functioning domain. Our data demonstrate that the burden of HAE on the quality of life of pediatric patients and their families encompasses a wide range of daily functioning.

The Prevalence of Atopic Dermatitis and Food Allergy in Children Living in an Urban Agglomeration-Is There a Current Relationship?

Atopic dermatitis (AD) prevalence in Poland is more frequent in individuals who live in a city. There are more studies demonstrating that long-term exposure to air pollutants is an independent risk factor for developing AD. The aim of the study was to assess the epidemiology of AD and food allergy (FA) in school children and adolescents living in Krakow, and to find a potential relationship between the incidence of atopic dermatitis with exposure to polluted air. In this paper, we presented the incidence of AD and FA between 2014 and 2018. We analyzed data collected from nearly 30,000 children aged 7-8 and adolescents aged 16-17 from the population of children and youth in Krakow. We correlated it with annual mean concentrations of PM10 and PM2.5, which indicated a gradual improvement in the air quality in Krakow. As our research results show that the prevalence of atopic dermatitis decreased with food allergy prevalence depending on the age group. We can suspect that this is the result of children growing out of a food allergy. It may be also influenced by more consequential eating habits in a group of adolescents and the elimination of allergenic foods from the diet. The decreasing incidence of atopic dermatitis appears to be also related to improvement in air quality.

Trends in the Epidemiology of Allergic Diseases of the Airways in Children Growing Up in an Urban Agglomeration.

The prevalence of asthma and allergies among children has become an increasing problem in the last few decades. Data on the population of children and adolescents, especially living in polluted cities, are limited and based on studies carried out in small groups. In our study, we analyzed the incidence of asthma and allergic rhinitis between 2014 and 2018. We analyzed data collected from nearly 30,000 children aged seven to eight and adolescents aged 16-17, which allowed us to assess the frequency of allergic diseases in the population of children and youth in Krakow. More than 40% of respondents reported allergic symptoms, and nearly 50% of them were not diagnosed and treated. In the group of seven- and eight-year-olds with a positive history of allergies, 52% had allergic rhinitis and 9.1% had asthma. In the group of 16-17-year-olds, allergic rhinitis was diagnosed in 35.8%, while asthma was found in 4.9% of cases. The results obtained over the course of 10 years has shown the reduction in the frequency of asthma (from 22% in the case of asthma in both age groups) and allergic rhinitis cases (from 63.9% in adolescents). In our opinion, this can be considered a positive effect of the preventive measures taken in Kraków after 2010. Although there is still a higher incidence of allergic diseases among children and young people living in urban areas compared to rural areas, the trend apparently has reversed for some diseases.

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.

The Influence of Air Pollution on the Development of Allergic Inflammation in the Airways in Krakow's Atopic and Non-Atopic Residents.

Until now, the simultaneous influence of air pollution assessed by measuring the objective marker of exposition (1-hydroxypirene, 1-OHP) and atopy on the development of allergic airway diseases has not been studied. The aim of this study was to determine the pathomechanism of the allergic response to PM2.5 in atopic and non-atopic patients. We investigated the changes in peripheral blood basophil activity of patients after stimulation with the birch pollen allergen alone, the allergen combined with PM2.5 (BP), PM2.5 alone, a concentration of 1-OHP in urine, and a distance of residence from the main road in 30 persons. Activation by dust alone was positive for all concentrations in 83% of atopic and 75% of non-atopic assays. In the group of people with atopy, the simultaneous activation of BP gave a higher percentage of active basophils compared to the sum of activation with dust and birch pollen alone (B + P) for all concentrations. The difference between BP and B + P was 117.5 (p = 0.02) at a PM concentration of 100 µg. Such a relationship was not observed in the control group. The correlation coefficient between the distance of residence from major roads and urinary 1-OHP was 0.62. A Pearson correlation analysis of quantitative variables was performed, and positive correlation results were obtained in the atopy group between BP and 1-OH-P. Exposure to birch pollen and PM2.5 has a synergistic effect in sensitized individuals. The higher the exposure to pollutants, the higher the synergistic basophil response to the allergen and PM in atopic patients.

Changes in qualitative and quantitative traits of birch (Betula pendula) pollen allergenic proteins in relation to the pollution contamination.

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.

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.

Bradykinin and oxidative stress in patients with hereditary angioedema due to C1 inhibitor deficiency.

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.

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.

Sputum biomarkers during aspirin desensitization in nonsteroidal anti-inflammatory drugs exacerbated respiratory disease.

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.

Hymenoptera sting in the head and neck region is not a risk factor for grade IV systemic reactions in patients with venom allergy.

INTRODUCTION Hymenoptera insect stings (ISs) in the head and neck (H&N) 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 H&N region were related to a higher risk of grade IV SRs in patients with HVA. PATIENTS AND METHODS This retrospective cross­sectional 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%; P <0.001 for age and P = 0.002 for sex). The IS site was divided into 7 categories. RESULTS The H&N region was the most common site for the IS (one­third 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 H&N region. The H&N 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 H&N region were not confirmed to be a risk factor for grade IV SRs in patients with HVA, regardless of age and sex.

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.

Antibody reactivity in patients with IgE-mediated wheat allergy to various subunits and fractions of gluten and non-gluten proteins from ω-gliadin-free wheat genotypes.

[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.

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.

Comparison of the basophil activation test versus the nasal provocation test in establishing eligibility for specific immunotherapy.

INTRODUCTION    Allergic rhinitis (AR) is the most common atopic disease. Specific immunotherapy (SIT) is the only effective treatment method for AR. In uncertain diagnostic cases, before establishing eligibility for SIT, nasal provocation tests (NPTs) should be performed. However, there are numerous contraindications to performing NPTs, and there is ongoing search for an alternative in vitro method. OBJECTIVES    The aim of the study was to determine whether a specific in vitro provocation, that is, the basophil activation test (BAT), may replace a specific in vivo provocation, that is, the NPT, in establishing patient's eligibility for SIT. PATIENTS AND METHODS    The study included 30 patients with AR caused by allergy to house dust mite or birch pollen, referred for SIT. The assessment of basophil activation by measuring CD63 antigen expression was performed using the Flow2 CAST test. Basophils were stimulated with allergen preparation (concentrations of 5000, 500, and 50 standardized biological units) used in NPTs. BAT results were expressed as stimulation index (SI) and basophil reactivity (BR). RESULTS    Allergen concentrations of 500 and 50 SBU proved to be appropriate for basophil stimulation. Median SI and BR were higher for positive NPT results than for negative NPT results (P <0.001). Sensitivity for SI and BR was in the range from 83% to 100%; specificity, from 78% to 89%; positive predictive value, from 75% to 87%; and negative predictive value, from 89% to 100%. We observed a high correlation of the analyzed parameters for the allergen concentrations of 500 and 50 SBU (range, 0.58-0.74; P <0.05). CONCLUSIONS    If there are contraindications to performing the NPT, BAT may be regarded as an alternative in establishing patients' eligibility for SIT. The optimal concentrations of allergen preparations are 500 and 50 SBU. Both SI and BR are good indicators of basophil activation.

[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.

[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.