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Hepatitis B nucleic acid vaccine and construction method thereof

A nucleic acid vaccine, hepatitis B technology, applied in the field of biomedicine, can solve the problems of unsatisfactory immune prevention effect and treatment effect, unsatisfactory effect, low immune reactivity, etc., and achieves good immune effect, long-lasting immune effect, The effect of simplifying the production process

Inactive Publication Date: 2011-01-26
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its immune prevention and treatment effects have been unsatisfactory, especially for chronic HBV-infected patients, due to the long-term presence of corresponding antigens in the body, the body's immune system has formed an immune tolerance state for HBV antigens, so after being inoculated with the vaccine, Immunoreactivity is much lower than normal
Therefore, the effect of hepatitis B DNA vaccine on immune tolerance and activation of HBV-specific T cell immunity in patients with chronic hepatitis B is not ideal

Method used

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  • Hepatitis B nucleic acid vaccine and construction method thereof
  • Hepatitis B nucleic acid vaccine and construction method thereof
  • Hepatitis B nucleic acid vaccine and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Construction of the nucleic acid vaccine of the present invention

[0031] The DNA vaccine is constructed by molecular biology experimental methods such as PCR, restriction digestion, ligation, transformation, etc., which can be referred to the second edition of "Molecular Cloning" written by Sambrook, USA. The PCR primer design software is Primer PREMER V from Premier Biosoft International.

[0032] The basic materials for constructing this vaccine include: human monocyte THP-1 (purchased from the Cell Bank of the Chinese Academy of Sciences), Hep G 2.215 liver cancer cells stably transfected with HBV whole virus (purchased from the Wuhan Institute of Virology, Chinese Academy of Sciences); eukaryotic expression vector pcDNA TM 3.1 / V5-His B plasmid (purchased from Invitrogen).

[0033] The reagents required for constructing this accounting virus include: Prime Star high-fidelity DNA polymerase (purchased from TaKaRa), pMD18T cloning vector (purchased from TaKaRa),...

Embodiment 2

[0108] Example 2: Animal experiment of the nucleic acid vaccine of the present invention

[0109] In order to verify the protective efficacy of the present invention against HBV infection, the present invention compared pcDNA3.1-SC-181a, pcDNA3.1-S expressing HBV S antigen, and pcDNA3.1-C expressing HBV C antigen and e antigen against mice. The difference in immune effect.

[0110] The experimental mice were female 8-week-old Balb / C mice, 30 of which were divided into 3 groups, 10 in each group. One group was injected with pcDNA3.1-S-C-181a, the second group was injected with pcDNA3.1-S, and the third group was injected with pcDNA3.1-C. The injection method was intramuscular injection, and the injection amount was 0.1 mg / mouse. In order to enhance the immune effect, insert the electric pulse shock vertically after the injection with the injection hole as the center. The electrode needle is inserted slightly deeper than the injection depth, the electric field strength is 200v / cm, t...

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Abstract

The invention relates to the technical field of biomedicines. The currently reported hepatitis B virus vaccine mainly comprises one or several of HBsAg, preS 1, preS2 and HBcAg, some cell factor genes with immunological enhancement function or lymphocyte epitope genes and the like, however, the immunoprophylaxis effect and treatment effect of the vaccine are always unsatisfactory. The invention provides a hepatitis B vaccine which comprises hepatitis B virus surface antigen gene HBsAg, core protein gene HBc and e-antigen gene, also comprises a human microRNA181a precursor gene sequence, and can assist stimulating the body immunity pathway. The construction process of the vaccine relates to PCR, enzyme cutting, connection, conversion and other molecularly biological operating means. The hepatitis B vaccine has the advantages of effectively activating the body to produce a specific antibody against the hepatitis B virus, stimulating body cell immunity and secreting various cell factors. Therefore, the aim of treating chronic hepatitis B is fulfilled.

Description

Technical field [0001] The invention belongs to the field of biomedical technology, and specifically relates to a new DNA vaccine against HBV virus and a construction method thereof. Background technique [0002] Hepatitis B is a worldwide infectious disease caused by hepatitis B virus (HBV). my country is an area with a high incidence of HBV infection. About 700 to 800 million people have been infected with hepatitis B, and about 120 million people are carriers of the hepatitis B virus. 1 / 4 of them develop chronic hepatitis, cirrhosis or primary liver cancer. Since I started to receive hepatitis B vaccine in 1985, the number of hepatitis B virus carriers among immunized children has dropped from 10% to about 1%. Therefore, practice has proved that hepatitis B vaccine is the most effective weapon to prevent and control hepatitis B. [0003] Hepatitis B virus is a hepatotropic DNA virus with a double shell structure. The outer shell is composed of hepatitis B virus surface antigen...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N15/51C12N15/79A61P1/16A61P31/20
Inventor 孙树汉魏伟章意亮商京丽
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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