Cockroach Allergen-Induced Asthma Model

The cockroach allergen–induced asthma mouse model was established with the protocol as illustrated in Figure 1A. Briefly, mice were sensitized by intra-tracheal (i.t.) inhalation of 20 µg of cockroach extract (CRE, B46, GREER Laboratories) per mouse in 50 μl of Phosphate-Buffered Saline (PBS) under light anesthesia with isoflurane and challenged with the same amount of CRE. Control mice were treated with PBS. Mice were sacrificed, and samples [lung tissues, bronchoalveolar lavage fluid (BALF), and blood] were collected on the next day after the last allergen challenge (30 (link)). In particular, BALF was harvested by two consecutive flushes of the lung with 1.0 ml of ice-cold PBS. Blood was taken to screen for serum antibodies against cockroach allergen. In some cases, mice were pre-treated with autophagy inhibitor chloroquine (CLQ; C6628, 60 mg/kg, Sigma-Aldrich, St. Louis, MO, USA) or saline vehicle control by intraperitoneal administration 1 h before every single allergen challenge.

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Wang J., Zhao Y., Zhang X., Tu W., Wan R., Shen Y., Zhang Y., Trivedi R, & Gao P. (2022). Type II alveolar epithelial cell aryl hydrocarbon receptor protects against allergic airway inflammation through controlling cell autophagy. Frontiers in Immunology, 13, 964575.

Publication 2022

C6628

Allergen Anesthesia Antibodies Asthma Autophagy Blood Bronchoalveolar lavage fluid Chloroquine Cockroach Cold Flushes Inhalation Intraperitoneal administration Isoflurane Light Lung Mice Phosphate Saline Serum Tissues Tracheal

Corresponding Organization :

Other organizations : Johns Hopkins University, Johns Hopkins Medicine, Sichuan University, West China Hospital of Sichuan University, Air Force Medical University, Xijing Hospital, Shenzhen Third People’s Hospital, Shenzhen University, Xiangya Hospital Central South University, Central South University

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Variable analysis

independent variables

Intra-tracheal (i.t.) inhalation of 20 µg of cockroach extract (CRE) per mouse
Administration of chloroquine (CLQ; 60 mg/kg) or saline vehicle control by intraperitoneal injection 1 h before every single allergen challenge

dependent variables

Lung tissues
Bronchoalveolar lavage fluid (BALF)
Blood (to screen for serum antibodies against cockroach allergen)

control variables

Control mice treated with phosphate-buffered saline (PBS) instead of cockroach extract (CRE)

controls

Positive control: Mice treated with cockroach extract (CRE)
Negative control: Mice treated with phosphate-buffered saline (PBS)

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