alias: GSM4546503
title: GSM4546503: Pat2_cHCV_P1_3; Homo sapiens; RNA-Seq
library_construction_protocol: PBMCs from EDTA anticoagulated patient blood were isolated by density gradient centrifugation using Pancoll (Pan-Biotech, Germany). Magnetic bead-based enrichment of antigen-specific CD8 T cells was further performed. For magnetic bead-based enrichment of HCV-specific CD8+ T cells PBMCs were incubated with peptide-loaded HLA- tetramers coupled to phycoerythrin (PE). Subsequent enrichment was performed with anti-PE beads using the MACS technology (Miltenyi Biotec, Germany) according to the manufacturer's protocol. Enriched HCV-specific CD8+ T cells were used for multiparametric flow cytometry and transcriptome analysis. Epitope-specific HLA tetramers were generated by conjugation of biotinylated peptide-MHC class I monomers with PE-conjugated streptavidin at a HLA:Strepatividin molar ratio of 5:1. The following peptide-loaded HLA tetramers were used: NS31073(CINGVCWTV)/HLA-A*02:01; NS31406(KLVALGINAV/KLSGLGLNAV)/HLA-A*02:01; NS52594(ALYDVVTKL)/HLA-A*02:01; and NS5B2841(ARMILMTHF)/HLA-B*27:05. As described in mCEL-Seq2 protocol (Hashimshony et al. 2016 and Herman et al. 2018) Adapted from TruSeq Small RNA Library Preparation Protocol
identifiers.namespace: GEO
identifiers.id: GSM4546503