Allergen
H.scandens pollen was purchased from
Beijing Key Laboratory of Precision Medicine for Diagnosis and Treatment on Allergic Diseases
(Beijing, China). HSE was prepared using the extraction method
previously described for Artemisia vulgarispollen[17]. Briefly,H.scandens pollen was defatted with acetone, extracted with 0.125
M ammonium bicarbonate(weight/volume=1:20), dialyzed against distilled
water, then aliquoted into vials (2.15 mL/vial) and freeze-dried.
Subsequently, freeze-dried HSE was diluted in
phosphate-buffered saline
(PBS) before use. The presence of
endotoxin in the pollen and HSE was assessed using the limulus amebocyte
lysate test, and the test results were qualified. The total protein
concentration was determined by the Bradford method. The concentration
of HSE dissolved in 200 μL PBS was 5.6
μg/μL (total protein, 1.12 mg/vial).
Experimental design