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An anti-hcv vaccine and preparation methods and uses thereof

A vaccine and purpose technology, applied in the field of genetic engineering, can solve the problems of damage to DC, narrow scope of applicable population, weak immune response, etc.

Inactive Publication Date: 2013-01-02
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The immune response induced by HCV peptide vaccine, which has been studied more at present, is weak, and is limited by HLA type, and the scope of applicable population is narrow, and the expression of HCV structural gene has been proved to damage the function of DC. Therefore, HCV non-structural gene should be a Main Candidate Genes for Preparation of HCV Vaccine

Method used

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  • An anti-hcv vaccine and preparation methods and uses thereof
  • An anti-hcv vaccine and preparation methods and uses thereof
  • An anti-hcv vaccine and preparation methods and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Construction of Adenoviral Vectors Carrying Various Nonstructural Genes of HCV and Expression Identification of HCV Proteins

[0033] 1. Experimental materials

[0034] Sera from HCV-infected patients were obtained from voluntary donors with signed informed consent.

[0035] Pshuttle-CMV (Germany, Merck, Cat. No. ST240007), Adeasy-1 plasmid (Germany, Merck, Cat. No. ST240005), Escherichia coli BJ5183 and recombinant adenovirus carrying GFP (AdGFP) were purchased from Beijing Nuosai Genome Research Center limited company;

[0036] Huh7 liver cancer cell line was purchased from People's Hospital (Jia Yintang et al., Construction of eukaryotic expression vector of interferon-stimulated gene ISG20 and its anti-hepatitis C virus replication, Chinese Journal of Immunology, 2006Vol.22No.11P.997-1001) ;

[0037] DH5α chemical transformation competence was purchased from Beijing Dingguo Biotechnology Co., Ltd.;

[0038] M-MLV Rtase cDNA synthesis Kit and LA-Taq DN...

Embodiment 2

[0121] Embodiment 2: healthy human body experiment in vitro

[0122] 1. Experimental materials

[0123] 1) Research object: The peripheral blood of 19 HLA-A2 positive healthy people was obtained from Beijing Red Cross Blood Center.

[0124] 2) Lymphocyte stratification fluid: purchased from Dingguo Biotechnology Co., Ltd., specific gravity 1.077g / ml; recombinant human granulocyte-macrophage colony-stimulating factor (rh GM-CSF) and recombinant human interleukin 4 (rh-IL -4) Purchased from an American R&D company;

[0125] AIM-V serum-free medium was purchased from GIBCO, USA;

[0126] RPMI 1640 medium was purchased from Dingguo Biotechnology Co., Ltd. Complete RPMI 1640 contained 10% fetal bovine serum, 1% glutamine, 100IU / ml streptomycin and 100μg / ml penicillin;

[0127] Mouse anti-human HLA-A2 monoclonal antibody was purchased from BD Company in the United States, and LPS and mitomycin were purchased from Sigma Company in the United States;

[0128] Hhu7 cells (HCVR) sta...

Embodiment 3

[0170] Embodiment 3: In vitro experiment of HCV infected persons

[0171] 1. Experimental materials

[0172] Cit

[0173] 2. Experimental method

[0174] The treatment method of the HCV infection group was the same as that of the D group in Example 2. When non-adherent PBMCs interacted with DCs, they first interacted for 24 hours, added IL-230IU / ml, replaced half of the medium every other day, and continued to cultivate for 48 hours. Others are the same as before.

[0175] 3. Experimental results (see Table 7)

[0176] Adenoviral vectors carrying HCVNS3 / NS4 and HCVNS4 / NS5 genes respectively induced strong cellular immune responses, but there was no statistical difference between them, and both were stronger than the immune responses induced by DCs loaded with 7 peptides.

[0177] Table 7HCV infection group IFN-γ, IL-4 and GrB ELISPOT results (SFC / 2×10 5 PBMC)

[0178]

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Abstract

An anti-HCV vaccine, which is prepared through recombination of an NS gene in series, including NS3 / NS4 and NS4 / NS5, with an adenovirus vector. The preparation methods and uses of the anti-HCV vaccine.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a vaccine against hepatitis C virus, a preparation method and application of the vaccine. Background technique [0002] Hepatitis C virus (HCV) infection is an important public health problem. There are 170 million HCV infections in the world, and the standardized prevalence rate of HCV infection in my country is about 3.2%. The notable feature of HCV infection is the high rate of chronicity, and chronic HCV infection is the main cause of end-stage liver diseases such as liver cirrhosis and hepatocellular carcinoma. [0003] Clearing the virus can improve liver inflammation, inhibit liver fibrosis and prevent disease progression. Interferon (IFN) α combined with ribavirin (RBV) is the only relatively effective drug for the treatment of HCV infection, but the sustained virological response rate (sustained virological response, SVR) is low, and there are still some p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/51C12N15/85C12N15/86C12N15/861C07K14/18A61K48/00A61P1/16A61P31/14
CPCA61K2039/5256C12N2710/10343A61K2039/53A61K39/29C07K14/005C12N2770/24222C12N2770/24234A61K39/12A61P1/16A61P31/14
Inventor 魏来田园
Owner PEOPLES HOSPITAL PEKING UNIV
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