Fig. 2.

Ex vivo induction of IFN-λs in PBMC from healthy volunteers. a mRNA expression levels of IFN-λs by real-time quantitative PCR. After pre-treatment with or without 100 U/ml of IFN-α for 16 h, 100,000 of mononuclear cells were stimulated with 30 μg/ml of poly I:C (a TLR3 agonist) or 5 μg/ml of R-837 (a TLR7 agonist). After stimulation with TRL-agonists for 4 h, the PBMC were harvested. b Protein levels of IFN-λs. After pre-treatment with 100 U/ml of IFN-α, 200,000 of mononuclear cells were stimulated with 30 μg/ml of poly I:C (a TLR3 agonist) or 5 μg/ml of R-837 (a TLR7 agonist), and the supernatant was harvested 24 h after stimulation with TLR agonists. c Kinetics of IFN-λs mRNA levels. After pre-incubation with IFN-α for 16 h, PBMC was stimulated with 5 μg/ml of R-837. Real-time quantitative PCR was conducted at each time point. d CLEIA results for IFN-λ3 protein in the supernatant at each time point