Ifn γ

The excised lungs were weighed, washed and homogenized in PBS. The expression of cytokines and chemokines,including TNF-α, IFN-γ, IL-10, IL-1β, MCP-1, IL-6, IFN-α (Neobioscience, Beijing, China), and CXCL1 (R&D Systems),was assayed in the supernatants from lung homogenates and BAL fluid using ELISA. The manufacturer's recommendations were followed during the assay.

Anti-PE MicroBeads were purchased from Miltenyi (Germany). Th1/Th2/Th17 phenotyping kit, FITC Hamster anti-Mouse TCR- (T cell receptor) β chain, PerCP-Cy™5.5 Mouse anti-T-bet (T box expressed in T cells), PerCP-Cy™5.5 Mouse anti-ROR-γt, Alexa Fluor 647 Mouse anti-PLZF (promyelocytic leukemia zinc finger protein), Th1/Th2/Th17 cytokines kit, DiR (1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbocyaine iodide) were purchased from Thermo Fisher Scientific (USA). PE-labeled T-selected-CD1d Tetramer was purchased from MBL International (Japan). KRN7000 (α-GalCer) was purchased from AdipoGen. Mouse lymphocyte isolate was purchased from Solarbio (Beijing, China). The MS Columns sorting column was purchased from Miltenyi (Germany). The small animal in vivo imaging system was purchased from PerkinElmer (USA). Phorbol 12-myristate 13-acetate (PMA) and ionomycin (IO) were purchased from Cayman company (USA). Mouse IL-17A, TNF-α, IFN-γ, and IL-4 cytokine ELISA Kit were purchased from Neobioscience Technology Co, Ltd. (Shenzhen, China).

The concentrations of IFN-γ, IL-4, IL-6, IL-10, and IL-17 (Neobioscience, Shenzhen, China) in the spleen CD4⁺ T cell culture supernatants or serum of mice were determined by commercially available ELISA kit following the manufacturer's instructions.

Serum levels of various cytokines were measured using ELISA kits according to the manufacturer’s instructions. The cytokines tested included IFN-γ, TNF-α, and IL-17 (Neobioscience Technology, Shenzhen, China) and KLK12, MMP-1, and MMP-9 (Jonln, Shanghai, China).