Sensitization and antigen challenge with OVA were performed as previously described.17 (link)18 (link) Mice were immunized by subcutaneously injecting 25 µg of OVA (Grade V; Sigma-Aldrich, St. Louis, MO, USA) absorbed to 1 mg of aluminum hydroxide (Aldrich, Milwaukee, WI, USA) in 200 µL of phosphate-buffered saline (PBS). Subcutaneous injections were administered on days 0, 7, 14, and 21, followed by intranasal OVA challenge (20/50 µL in PBS) performed on days 33 and 35. Subsequently, intranasal OVA challenges were repeated twice per week for 3 months. Age- and gender-matched control mice were treated equally with PBS. All procedures were performed while mice were anesthetized using isoflurane (Vedco, St. Joseph, MO, USA). Mice were sacrificed 24 hours after the final intranasal OVA challenge, and bronchoalveolar lavage (BAL) fluid and lung tissues were obtained for analysis.
All animal procedures were performed in accordance with Laboratory Animal Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Rodent Experiments provided by Institutional Animal Care and Use Committee at the School of Medicine, The Catholic University of Korea (apprOVAl number: CUMC-2015-0194-04).
All animal procedures were performed in accordance with Laboratory Animal Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Rodent Experiments provided by Institutional Animal Care and Use Committee at the School of Medicine, The Catholic University of Korea (apprOVAl number: CUMC-2015-0194-04).
