Construction of hepatitis C virus CTL-Th epitopes chimeric DNA vaccine
1.Department of Liver, the Second Hospital of Ningbo, Ningbo, 315000; 2.Institute of Arboviruses, Wenzhou Medical University, Wenzhou, 325035; 3.Department of Infection, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015
LI Dezhou1,DUAN Zhiliang2,GUO Jianglong2, et al. Construction of hepatitis C virus CTL-Th epitopes chimeric DNA vaccine[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2015, 45(2): 79-.
Abstract：Objective: To construct hepatitis C virus (HCV) CTL-Th epitopes chimeric DNA vaccine and explore its in vivo immune response. Methods: Based on previously identified four HCV-specific HLA-A*0201-restricted CTL (Cytotoxic T Lymphocyte) epitopes (NS4b_78, NS5a_367, C_181 and NS2_172), two HLA-A*1101-restricted CTL epitopes (NS3_609 and NS5b_251), one HLA-A*2402-restricted CTL epitopes (NS5b_382) and two Th epitopes (P73-15 and NS5A276-288), the gene encoding chimeric CTL epitopes and Th epitopes was synthesized and cloned into eukaryotic expressing vector pcDNATM3.1/myc-His(-) A. The recombinant plasmid was used to immunize HLA-A*0201, HLA-A*1101 and HLA-A*2402 transgenic mice, ELISPOT and CTL cytotoxicity assay were used to measure the frequencies of CTL epitope-specific T cells in splenocytes of mice and the cytotoxic activity of CTL against target cells. Results: The gene containing Kozak sequence and encoding Igκ chain signal sequence, seven CTL epitopes, two Th epitopes was successfully cloned into eukaryotic expressing vector. Recombinant plasmid immunization elicited epitope-specific IFN-γ-secreting T cells which could lyse CTL epitope-pulsed splenocytes. Conclusion: A Hepatitis C virus CTL-Th epitopes chimeric DNA vaccine which can induce epitope-specific CTL response is successfully constructed.
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