Protocol for the vaccination and intranasal exposure to house dust mite.

Blood was collected on day -21 to detect the baseline of anti-IL-33 IgG level. Vaccine (100 μg of IL-33 recombinant protein or carrier protein) was administrated subcutaneously from day-20, three times at a two-week interval. On days 0 (one week prior to the final immunization), 1, 2, 5, 6, 7, 12, 13, 27, 28, 41 and 42, mice were challenged by intranasal administration of HDM extract (25 μg) in 20 μl PBS or equal volume PBS On day 44 lung physiology assessment was performed and samples were taken for analysis. HDM, house dust mite; Trx, thioredoxin; i.n., intranasal; PBS, phosphate buffered saline.