Bronchoalveolar lavage (BAL) samples were obtained as described previously [10 (link)]. Briefly, the trachea was exposed and intubated with a catheter, and 2 sequential lavages were performed in each mouse by injecting sterile PBS. The recovered fluid was centrifuged and frozen at −70 °C for subsequent analyses. Albumin content, a measure to quantitate increased permeability of the bronchoalveolar–capillary barrier, and lactate dehydrogenase (LDH) activity, an indicator of general cytotoxicity, were determined in the acellular BAL fluid as described previously [10 (link),18 (link)].
Interferon (IFN)-β (Mouse IFN-beta ELISA Kit, sensitivity: 15.5 pg/mL), IFN-γ (Mouse IFN-gamma Quantikine ELISA Kit, sensitivity: 2 pg/mL) and IL-10 (Mouse IL-10 Quantikine ELISA Kit, sensitivity: 5.2 pg/mL) concentrations in BAL samples were measured with commercially available enzyme-linked immunosorbent assay (elisa) technique kits following the manufacturer’s recommendations (R&D Systems, Minneapolis, MN, USA).
Interferon (IFN)-β (Mouse IFN-beta ELISA Kit, sensitivity: 15.5 pg/mL), IFN-γ (Mouse IFN-gamma Quantikine ELISA Kit, sensitivity: 2 pg/mL) and IL-10 (Mouse IL-10 Quantikine ELISA Kit, sensitivity: 5.2 pg/mL) concentrations in BAL samples were measured with commercially available enzyme-linked immunosorbent assay (elisa) technique kits following the manufacturer’s recommendations (R&D Systems, Minneapolis, MN, USA).
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