Airway Hyperresponsiveness in Gαi1/3 Knockout Mice

WT or Gαi1/3 DKO mice were sensitized by intraperitoneal injection of ovalbumin (OVA, twice, one week apart, Sigma) 32 (link). One week following the last sensitization, mice were anesthetized and challenged with OVA or PBS as described 32 (link). Airway responsiveness, pulmonary inflammation and immunoglobulin synthesis were compared in wild-type and Gαi1/3 DKO mice sensitized and challenged with PBS or OVA. Three days after aspiration challenge, airway responsiveness to intravenous acetylcholine chloride (Ach) administration was determined using the described protocol 32 (link). The number of inflammatory cells in bronchoalveolar Lavage (BAL) was determined. Lungs were also fixed and subjected to HE staining and Masson staining. Mouse lung tissues were digested and minced as reported 33 . After lysis of red blood cells (RBCs), the dissociated cells were underlaid with 7.5 mL of lymphocyte separation medium (Sigma, Shanghai, China) and cells were centrifuged. From the middle layer the mononuclear cells were incubated in six-well plates for two hours 33 . Thereafter, the adherent cells were alveolar macrophages.