Aluminum hydroxide adjuvant

Mouse models were established as previously described [8 (link)]. In brief, on Days 0, 7, and 14, the AS, EB, and DXM groups were administered with OVA (Sigma Company, USA, A5503-10G) sensitization solution at 200 μL/animal (containing 10 µg of OVA + 1.3 mg of aluminum hydroxide adjuvant (Thermo Fisher Scientific, USA, 77161)) by intraperitoneal injections and challenged with 200, 10, and 10 µg OVA on Days 21–23, respectively. Additionally, the DXM group was treated with 5 mg/kg of DXM (Guangzhou Baiyunshan Tianxin Pharmaceutical Co., Ltd., China, H44022091) by intraperitoneal injection 1 hr before stimulation and measurement of airway reactivity. The NS group was administered equal amount of NS for intraperitoneal sensitization and intranasal stimulation.
In the Sema3E treatment protocol (Figure 1(a)), the AS mouse model was established as described above while challenged i.n. with OVA (200 µg in 25 µL NS), mice were exposed intranasally to Sema3E (5 µg in 25 µL NS) (R&D Systems, Minneapolis, MN., USA, 3239-S3B-025), or NS as a control 1 hr before challenge. Twenty-four hours after the last administration, mice were anesthetized for invasive airway resistance detection and then sacrificed for analysis of airway inflammation, mucus production, and collagen deposition.

Ginsenoside Rb1 was purchased from Beijing Solarbio Technology Co., Ltd. (Beijing, China) with product number SA9790 and purity ≥ 98%. Influenza A virus antigen and IIV4 were purchased from Zhejiang Tianyuan Biopharmaceutical Co., Ltd; the A/Michigan/45/2015 virus was stored in our laboratory. Aluminum hydroxide adjuvant was purchased from Thermo Fisher Scientific Co., Ltd (Waltham, MA, USA). Secondary antibodies were purchased from Southern Biotech. The cytokine kit was purchased from Beijing 4A Biotech Co., Ltd.