Earticle

초록

영어

N-linked glycosylation of viral proteins have been implicated in shielding antigen epitopes and blocking neutralizing antibodies production and contribute to the evasion of HCV from the host immune response. In this study, the effects of N-linked glycosylation of Hepatitis C virus (HCV) envelope E1 and E2 proteins, the naturally poor immunogens, on the induction of specific immune response were examined. We constructed plasmids containing the genes encoding both wild type E1E2 proteins and mutated E1E2 proteins in which N-linked glycosylation sites are mutated individually or in combination by site-directed mutagenesis or coupled with CpG, a TLR9 ligand. The immunoreactivity of wild type E1E2 and mutated E1E2 proteins or coupled with immune activator CpG was analyzed in BALB/C mice using a DNA-based vaccination approach. We found that E1E2 specific N-glycosylated mutant induced much higher IgG titer than wild type (WT) E1E2. E1E2 mutant vaccinated mice developed a mixture of IgG1 and IgG2a, but CpG-E1E2 mutant and E1-E2 wild type dominantly developed IgG2a isotype. And CpG-E1E2 mutant significantly stimulated Th1-type response and specific CD8+T cells cytotoxic T lymphocytes (CTL) activities. More important we found that CpG-E1E2 mutant significantly induced much higher neutalizing antibody than E1-E2-WT and had the highest level of neutralizing antibodies among all groups. The monoclonal clonal antibody (McAb) were prepared from CpG-E1E2 mutant immunized mice and the main neutralizing McAb 1C2 had significantly neutralizing Ab activity with blocking HCVcc infection in Huh7.5.1 cells in vitro. Epitope mapping of binding of a neutralizing McAb 1C2 was also analyzed. Our data indicate that the presence of glycans at the surface of HCV envelope proteins could help explain how HCV evades the humoral immune response and why most HCV infections lead to chronicity. CpG have the ability to augment both neutralizing antibody and cellular responses of E1E2 mutant. The CpG-E1E2 mutant holds potential applicant for the development of therapeutic vaccine with enhanced cellular and neutralizing immune responses.

저자

• Yushan Ren [ State Key Laboratory of Virology, Department of Immunology and Hubei province Key Laboratory of Allergy and Immune-related diseases, Wuhan University School of Medicine, Wuhan 430072, China ]
• Miao Lin [ State Key Laboratory of Virology, Department of Immunology and Hubei province Key Laboratory of Allergy and Immune-related diseases, Wuhan University School of Medicine, Wuhan 430072, China ]
• Yuanqin Min [ State Key Laboratory of Virology, Department of Immunology and Hubei province Key Laboratory of Allergy and Immune-related diseases, Wuhan University School of Medicine, Wuhan 430072, China ]
• Xiao-Lian Zhang [ State Key Laboratory of Virology, Department of Immunology and Hubei province Key Laboratory of Allergy and Immune-related diseases, Wuhan University School of Medicine, Wuhan 430072, China ] Corresponding author

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발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614