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Objective: To perform a genome-wide characterization of changes in microRNA (miRNA) expression during the course of venom immunotherapy (VIT)., Methods: miRNA was isolated from the whole-blood of 13 allergic patients and 14 controls, who experienced no allergic reaction upon stings by honeybees and wasps. We analyzed 2549 miRNAs from the whole blood of these patients prior to VIT and 12 months after the start of VIT. The results for differential expression obtained on a microarray platform were confirmed by quantitative real-time PCR. Out of the 13 patients, 8 had a negative allergic reaction with VIT, thus indicating that this approach was successful., Results: By comparing time points before and 12 months after ultrarush VIT, correlation analysis and principal component analysis both indicated a limited effect of VIT on the overall miRNA expression pattern. Volcano plot analysis based on raw P values revealed few deregulated miRNAs, most of which were increasingly expressed after VIT as compared with before VIT. Based on the 50 most altered miRNAs, no clear clustering was observed before or after VIT., Conclusions: Our results indicate an overall reduced effect of VIT on the miRNA expression pattern in whole blood.

Objective: The aim of our study was to determine the diagnostic usefulness of a newly developed basophil activation test (BAT) in patients allergic to Dermatophagoides pteronyssinus and pollens. We also analyzed the influence of cetirizine on CD63 upregulation. This popular antihistamine strongly inhibits skin tests, but its impact on BAT sensitivity remains unknown and deserves at least preliminary determination., Methods: The study sample comprised 22 patients allergic to house dust mite and pollens and 19 healthy controls. All participants underwent skin prick testing and the newly developed flow-cytometric basophil activation test. The protocol for allergen-induced basophil CD63 upregulation consisted of whole blood samples that were processed and stained with anti-CCR3/CD63 antibodies added to the buffer at the beginning of stimulation. Skin prick tests and BAT were performed twice--before and 2 hours after ingestion of 10 mg of cetirizine., Results: The new BAT is characterized by its short processing time, easy basophil gating, and strong CD63 upregulation with very high sensitivity and excellent specificity. Our results suggest that allergen-induced CD63 upregulation by higher doses of allergens is not inhibited 2 hours after administration of cetirizine (unlike skin prick tests)., Conclusion: The BAT is a very useful and precise method for the diagnosis of allergy to aeroallergens. It is not influenced by cetirizine.

We present a case of anaphylactic shock induced by exercise following celery ingestion. The possible mechanism of food-dependent exercise-induced anaphylaxis (FDEIA) and the laboratory tests for its diagnosis are discussed. We evaluated spontaneous, celery-allergen-induced, and anti-FcepsilonRI-antibody-induced histamine release from basophils obtained from the patient, 2 celery-allergic controls, and 3 healthy controls. Buffers of increasing osmolarity were used to mimic conditions of vigorous physical exercise. Only the patient's basophils showed an increase in spontaneous, anti-FcepsilonRI antibody-induced and allergen-induced histamine release under physiological conditions and with slightly increased medium osmolarity. To our knowledge, this is the first report on the possible role of increased histamine releasability in the pathogenic mechanism of FDEIA. We suggest that FDEIA results from increased histamine releasability triggered by physical effort after exposure to a sensitizing food allergen.

Background: Airway remodeling has recently emerged as a major problem in an increasing percentage of patients with asthma. Reasons for great diversity in the progression of irreversible bronchoconstriction among asthmatics remain unclear., Objective: The aim of this study was to assess whether the potential ability of leukocytes to produce cysteinyl leukotrienes in response to various stimuli is correlated with magnitude of irreversible airway obstruction in asthmatics., Materials and Methods: The study sample comprised 76 asthmatics (34 males, mean +/- SD age 52 +/- 13 years), and 35 healthy controls (18 males, 38.2 +/- 15 years). Each subject underwent 2 pulmonary function tests: before and after bronchodilator administration. In addition, approximate annual decline in forced expiratory volume in 1 second (FEV1) (% of predicted) was calculated. Leukotriene C4 (LTC4) production was assessed combining a cellular antigen stimulation test and enzyme-linked immunosorbent assay using Bulhmann Laboratories AG kits. Leukocytes isolated from peripheral blood were stimulated with anti-FcepsilonRI antibody, N-formyl-methionyl-leucyl-phenylalanine (fMLP) and phorbol 12-myristate 13-acetate (PMA). In separate tubes each subject's leukocytes were tested for spontaneous LTC4 production. Finally, stimulated LTC4 production was expressed in pg/mL after subtraction of values of spontaneous production., Results: In asthmatics, baseline FVC% and FEV% values ranged from 24.4% to 122.4% (mean, 75.5%) and from 23.4% to 126.6% (mean, 74.4%), respectively. There were no significant differences between asthmatics and controls in LTC4 production stimulated by anti-FcepsilonRI antibody (P = .79), fMLP (P = .33) or PMA (P = .86). We found no correlation between stimulated LTC4 production and spirometric parameters at baseline or after bronchodilator administration or annual decline in FEV1%., Conclusion: Our results do not confirm the hypothesis that airway remodeling in asthma might be related to enhanced ability of leukocytes to produce cysteinyl leukotrienes in response to various stimuli.

Airway fungal infections are often associated in asthmatics with the exacerbation of asthma symptoms. However, the pathomechanism of this phenomenon has not been fully understood. The aim of our study was to assess whether antimycotic treatment can influence the capacity of bronchoalveolar (BAL) leukocytes to release proinflammatory cytokines, which could contribute to increase in asthma severity. Ten patients with bronchial asthma complicated by airway fungal infections (Candida albicans and/or Aspergillus fumigatus) were included in the study. Seven asthmatics were treated with systemic and inhaled corticosteroids, whereas the remaining three with inhaled ones only. All subjects underwent several courses of therapy with antibiotics due to respiratory infections. BAL leukocytes obtained from the patients were cultured in the absence or presence of lipopolysaccharide E.coli (LPS) or Newcastle disease virus (NDV). The BAL procedure and measurement of the levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (II-6), interferon-gamma (IFN-gamma), and interferon-alpha (IFN-alpha) by specific bioassays were performed twice: before antimycotic treatment and after 3 weeks of therapy with 8 mg of nebulized fluoconazole and 400 mg of oral ketoconazole per day. The elimination of fungi from respiratory tract resulted in an apparent clinical improvement. This coincided with diminished production of TNF-alpha in response to LPS and the production of IFN-alpha in response to NDV, which were initially high and subsided significantly after antimycotic therapy (p = 0.035, and 0.011, respectively). Such changes were not observed in the case of IFN-gamma and IL-6. This may suggest that TNF-alpha as well as IFN-alpha are secreted by fungi-prestimulated leukocytes from the lower respiratory tract and may be involved in the processes of exacerbation of asthma complicated by fungal infections. Further analyses of relationships between changes in cytokine levels and clinical parameters indicated that IFN-alpha seems to be of particular interest in fungal stimulation of asthma.