MNCs (5 × 105 cells) of liver or splenic origin in 200 μL of medium were cultured with either α-GalCer (100 ng/mL), ConA (1 ng/mL, Vector Laboratories, Inc., Burlingame, CA), anti-CD3 Ab (10 μg/mL, eBioscience), CpG-ODN (20 μg/mL, Hycult Biotech, Uden, The Netherlands), or LPS from E. coli 0111:B4 (10 μg/mL, Sigma-Aldrich, St. Louis, MO) in 96-well flat-bottom plates. The culture supernatants were harvested 48 hours after seeding.