Huh 7.5 cells

Naïve CD8⁺ T cells from the healthy controls and patients were isolated with a Naïve CD8⁺ T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). The cells were washed with RPMI 1640 and then used for co-culture, stimulation, and a flow-cytometric analysis. Naïve CD8⁺ T cells, PBMCs, and/or Huh 7.5 cells (American Type Culture Collection, Manassas, VA, USA) were co-cultured with Dulbecco’s modified Eagle’s medium (Mediatech Inc., Manassas, VA, USA) containing 2% foetal bovine serum (Life Technologies, Grand Island, NY, USA), 0.05 mM β-mercaptoethanol, 2 M l-glutamine (Mediatech Inc.), and 100 µg/ml penicillin–streptomycin. JFH/Huh 7.5 cells were produced by transfecting cells with the FL-J6/JFH chimeric strain of HCV. Briefly, HCV FL-J6/JFH mRNA was transcribed with a TranscriptAid T7 High Yield Transcription Kit (Fermentas, Vilnius, Lithuania), as previously described.^{31 (link)} Huh7.5 cells were transfected with the transcribed mRNA using DMRIE-C Reagent (Invitrogen). The transfected Huh7.5 cells were collected after 24 h for further experiments. The transfection efficiency was verified with an anti-NS5 antibody (BD Bioscience), as shown in Supplementary Figure 1(a).