Figure 1

Proliferation in response to peptides of wild-type NS3 _358-375 and its variants and binding affinity of those peptides to HLA-DRB1*1501 molecules. A and B: PBMC from a HCV-infected patient B3019 were cultured with wild type NS3_358-375 and variant peptides at the given concentrations, to six days, pulsed overnight with 1.0 μCi/well tritiated thymidine. Radioactive label incorporation (cpm) was measured. PBMC with tissue culture medium alone was used as a negative control. Experiments were performed in triplicate. A representative of five experiments is shown. C and D: Binding of biotinylated peptides to HLA-DRB1*1501 molecules were assessed using an antigen-capture ELISA. Overnight binding affinities of variant peptides were compared to wild type peptide NS3_358-375. A representative of three experiments is shown. E and F: PBMCs were pre-pulsed with 0.1 μM variant peptides for three hours, then re-challenged with wild type NS3_358-375 at a concentration of 10 μM. Rest of the experiment was as same as mentioned in A. A representative of five experiments is shown.