4.7. Murine cytokine profiling

Frozen serum samples and snap‐frozen lung homogenates from the right lower and middle lung lobes were assessed. Lung samples were thawed, weighted and homogenized on ice with lysis buffer containing 0.05% sodium azide, 0.5% Triton X‐100, 1:500 Protease inhibitor cocktail (all from Sigma‐Aldrich) in sterile PBS, using 1 ml per 100 mg of tissue. Homogenates were incubated for 1 h at 4°C and centrifuged at 3000 × g for 10 min. Supernatants were collected and stored at ‐80°C until use. Cytokines were measured by Luminex xMAP technology and analyzed with xPONENT 3.1 software (Luminex Corporation). IFN‐γ, TNF‐α, IL‐6, LIX and IL‐17 were measured in serum samples, using the MCYTOMAG‐70K kit. In lung homogenates, IFN‐γ, TNF‐α, IL‐4, IL‐6, LIX, IL‐1β, IL‐10, IL‐12, IP‐10, KC, MCP‐1 and VEGF were analyzed with the MCYTOMAG‐70K kit, and TGFβ with the TGFBMAG‐64K kit (EMD Millipore Corporation) following the manufacturer's instructions.