Selected article for: "PBMCs ml and RPMI medium"
Author: Kammer, Andreas R.; van der Burg, Sjoerd H.; Grabscheid, Benno; Hunziker, Isabelle P.; Kwappenberg, Kitty M.C.; Reichen, Jürg; Melief, Cornelis J.M.; Cerny, Andreas
Title: Molecular Mimicry of Human Cytochrome P450 by Hepatitis C Virus at the Level of Cytotoxic T Cell Recognition
  • Document date: 1999_7_19
    • ID: 1vabzhs5_7
    Snippet: Induction of Primary CTLs. PBMCs from HLA-A2-positive healthy donors and HCV patients were isolated on Ficoll-Paque density gradients and washed three times in PBS containing 10% FCS-medium. 4 Ï« 10 6 PBMCs were incubated with synthetic peptide (10 g/ml; Chiron Mimotopes) in RPMI 1640 medium supplemented with l -glutamine (2 mM), penicillin (50 U/ml), streptomycin (50 g/ml), and Hepes (5 mM) containing 10% (vol/vol) heat-inactivated human AB seru.....
    Document: Induction of Primary CTLs. PBMCs from HLA-A2-positive healthy donors and HCV patients were isolated on Ficoll-Paque density gradients and washed three times in PBS containing 10% FCS-medium. 4 Ï« 10 6 PBMCs were incubated with synthetic peptide (10 g/ml; Chiron Mimotopes) in RPMI 1640 medium supplemented with l -glutamine (2 mM), penicillin (50 U/ml), streptomycin (50 g/ml), and Hepes (5 mM) containing 10% (vol/vol) heat-inactivated human AB serum (AB-medium) for 1 h, washed once in PBS containing 10% FCS-medium, and then plated in 24-well plates at 4 Ï« 10 6 cells/well in AB-medium. On day 3 and weekly thereafter, 1 ml of complete medium supplemented with rIL-2 (20 U/ml; EuroCetus B.V.) was added to each well. On day 7 and weekly thereafter, the cultures were restimulated with 10 6 peptide-pulsed, irradiated (7,400 rads) autologous feeder cells in 1 ml of AB-medium containing rIL-2 (20 U/ml). The cultured PBMCs were tested for CTL activity against different peptides on day 35 .

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