The syringe with the pack and no piston was placed in a Falcon tube. AR sensitized to based on either positive results of pores and skin prick test (A/H percentage >3) or elevated serum specific IgE (>0.35 kU/L). The study individuals were divided into 2 organizations according to the NPT results: the NPT-positive (n=39) and NPT-negative (n=21) organizations. The individuals stopped receiving antihistamines, leukotriene receptor antagonists, or intranasal steroids prior to NPT for at least 3 days. The results of pores and skin prick test, levels of serum total/specific IgE to and value(%)38/39 (97.4)17/21 (80.1)0.002AR vs AR with BA27/1218/30.218Serum total IgE* (IU/mL)370.18412.33478.001,095.680.585Serum specific IgE to was performed according to previously described methods.16,17,18 Patients were first placed at space temperature for 30 minutes to minimize the effects of daily-life stimuli. They underwent saline challenge checks to exclude nose hyperreactivity. NPT was performed by applying an 8-mm filter paper disk impregnated with an allergen remedy (5,000 BU/L, was defined as a 3 point or more increase in the total nose symptom score, which is the sum of rhinorrhea, nose itching, nose obstruction, and sneezing during the test. Sample collection and processing The cotton ball nose packing method suggested by Ghent University or college Hospital19 which was modified according to conditions was applied. In brief, a Falcon tube with the cotton ball (Wejae Sangsa, Chungju, Korea) was weighed. Cotton ball nose packs were placed on the middle turbinate for 5 minutes after NPT to collect nose secretions. The cotton ball was placed back to the Falcon tube and weighed again to obtain the volume of nose secretions. After 3 mL of normal saline were added to the pack and incubated at 4 for 2 hours, the pack was put in the shaft of a syringe 10 mL (Kovax-Syringe; Korea Vaccine Co. LTD, Ansan, Korea). The syringe with the pack and no piston was placed in a Falcon tube. The Falcon tube was centrifuged at 1,500 g for 10 minutes at 4. Aliquots of 500 L were prepared and stored at -70 for analysis. Measurement of local antibodies, inflammatory mediators, and inflammatory markers in nose secretion samples The levels of total IgE and specific IgE to and in the nose secretion were recognized by using ImmunoCAP? trans-Zeatin (ThermoFisher Scientific, Uppsala, Sweden). The protein concentrations were determined by the Bradford assay. The levels of specific IgE, IgA, and secretory IgA to in the nose secretion were measured by using enzyme-linked immunosorbent trans-Zeatin assay (ELISA) as previously explained.5,16 Rabbit polyclonal to IWS1 In brief, 96-well microtiter plates (Corning, Corning, NY, USA) were coated with (2 g/well, homemade antigen)20 trans-Zeatin and incubated overnight at 4. After washing, the plates were clogged with 10% fetal bovine serum (FBS) in phosphate buffered saline (PBS) and incubated for 2 hours. Fifty microliters of the nose secretion was incubated for 2 hours. Then, biotin-labeled goat anti-human IgE antibody (Vector Lab, Burlingame, CA, USA) at 1:1,000 dilution, biotinylated anti-human IgA (Sigma Co., St. Louis, MO, USA) at 1:2,000 dilution, and monoclonal anti-secretory IgA (Sigma Co.) at 1:2,000 dilution were added and incubated for 1 hour to detect specific IgE, IgA, and secretory IgA to test between the NPT-positive and NPT-negative organizations. Symptom scores were compared by using Wilcoxon’s signed-ranks test. Pearson’s correlation coefficients were applied to examine the correlation between local antibodies and inflammatory mediators. A value of <0.05 was considered statistically significant. RESULTS Characteristics of the study individuals There was no significant difference in age or sex between the NPT-positive and NPT-negative organizations. Twelve of the 39 individuals in the NPT-positive group and 3 of the 18 individuals in the NPT-negative group experienced bronchial asthma. The baseline total nose symptom scores were 11.599.49 in the trans-Zeatin NPT-positive group and 6.817.33 in the NPT-negative group (was significantly higher in the NPT-positive group than in the NPT-negative group. The medical characteristics of the individuals are summarized in Table 1. Assessment of local antibody levels in nose secretions between the NPT-positive and NPT-negative organizations Fig. 1 shows assessment of total IgE, specific IgE, IgA, and secretory IgA.