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The DNA vaccine of an anti-infection of hepatitis C virus

A DNA vaccine and virus infection technology, applied in the fields of molecular microbiology and infection immunity, can solve the problems of large side effects

Inactive Publication Date: 2007-04-11
WUHAN UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The classic treatment for HCV infection is only interferon-alpha or a combination of interferon-alpha and nuclear analogue virus wah [4] , but the therapy has major flaws and side effects [5]

Method used

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  • The DNA vaccine of an anti-infection of hepatitis C virus
  • The DNA vaccine of an anti-infection of hepatitis C virus
  • The DNA vaccine of an anti-infection of hepatitis C virus

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Embodiment Construction

[0019] The present invention utilizes the technique of molecular cloning to successfully construct a recombinant DNA vaccine expressing HCV envelope glycoprotein E1, and construct six N-glycosylation mutants (M1-M6) of E1 on this basis, and use them as DNA vaccine, to study the effect of glycosylation on the body's immune response. It was found that E1-M2 can improve the host-specific cellular immune response and can be used as a new type of effective DNA vaccine against the immune response of HCV virus.

[0020] The recombinant DNA vaccine of above-mentioned E1 is made by comprising the following steps:

[0021] (1) Preparation of the prokaryotic expression vector of HCV E1:

[0022] Firstly, the whole genome of HCV was used as a template to carry out PCR reaction to obtain the HCV E1 DNA fragment, and to construct the HCVE1 prokaryotic expression vector pGEX-KG-HCV-E1.

[0023] Then, the obtained pGEX-KG-HCV-E1 was sequenced after being identified correctly by enzyme diges...

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Abstract

The invention relates to preparation and application of DNA vaccine that can resist to the infection of hepatitis C virus. The invention constructed a recombinant DNA vaccine that can express HCV enveloped glycoprotein E1 with molecular cloning techniques, on this basis constructed six N-glycosylation mutants M1-M6 of the E1 and select N-glycosylation mutant M2 as DNA vaccine that resist to infection of hepatitis C virus and application in the preparation of DNA vaccine that prevent the HCV virus infection. The advantages of the invention are : experiments reveal that N-sugar chain of HCV E1 enveloped glycoprotein can limit the host's cellular immune responses, its glycosylation may shield the T and B cell epitope of virus, thereby enable the virus to evade immune recognition; with E1 glycosylate mutants M2 may be used as DNA vaccines that are HCV therapeutic and preventive and enhance the immunogenicity, besides preparation of mutants M2 E1 glycosylate is simple, convenient and effective, and has good application prospect.

Description

technical field [0001] The invention relates to the fields of molecular microbiology and infection immunity, in particular to the preparation and application of a DNA vaccine against hepatitis C virus infection. Background technique [0002] Hepatitis C virus (HCV) infection is one of the most important causes of chronic liver disease [1] , my country is also one of the areas with a high incidence of hepatitis C, and HCV infection accounts for about 3.2% of the total population. About 80% of patients with acute HCV infection will become chronic persistent infection [2] 20% of the patients will progress to liver cirrhosis after 20 to 30 years, and 1 to 5% of the patients will progress to hepatocellular carcinoma (HCC). HCV has distinct genetic and phenotypic differences [3] , and the hepatitis C virus can only infect humans and chimpanzees, which makes research on hepatitis C in trouble. The classic treatment for HCV infection is only interferon-alpha or a combination of ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P31/14A61P1/16C12N15/74C12N15/79
Inventor 章晓联刘敏
Owner WUHAN UNIV
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