For culture of mouse T cells, splenocytes were collected and treated with ACK lysis buffer (Lonza, Switzerland) to remove red blood cells. Naïve T cells were isolated with a plate-bound anti-mouse CD3 antibody at 5 μg·mL−1 (eBioscience, USA). Activated T cells were obtained by stimulating naïve T cells for 48 h with a soluble anti-mouse CD28 antibody at 2 μg·mL−1 (eBioscience, USA) in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated FBS, 2 mmol·L−1 L-glutamine, 55 μmol·L−1 2-mercaptoethanol, 10 mmol·L−1 hydroxyethyl piperazine ethanesulfonic acid (HEPES), 1 mmol·L−1 sodium pyruvate, 100 U·mL−1 penicillin, and 100 μg·mL−1 streptomycin (all from Invitrogen, USA). Th1 cells were obtained by treating activated T cells with 20 ng·mL−1 IL-2 (PeproTech, USA), 20 ng·mL−1 IL-12 (PeproTech, USA), and 10 μg·mL−1 anti-IL-4 blocking antibody (BioLegend, USA).

Mouse macrophages were isolated by seeding 2 × 106 femoral ANCs in 6-well plates with 20 ng·mL−1 macrophage-colony stimulating factor (M-CSF; PeproTech, USA) in DMEM supplemented with 15% heat-inactivated FBS, 100 U·mL−1 penicillin and 100 μg·mL−1 streptomycin (all from Invitrogen, USA). After 48 h, nonadherent cells were removed, and the adherent macrophages were cultured for another 48 h with 20 ng·mL−1 M-CSF in the presence or absence of 1 ng·mL−1 LPS (Millipore, USA).

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