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Vaccine for controlling persistent infection of hepatitis B virus

A technology of hepatitis B virus and persistent infection, applied in antiviral agents, medical preparations containing active ingredients, biochemical equipment and methods, etc., can solve the problem of high protein

Inactive Publication Date: 2013-04-10
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hansenula is rich in carbon sources, and its protein production is higher than that of traditional Saccharomyces cerevisiae, and Hansenula itself is not pathogenic. There is no report on the use of Hansenula cells to enhance HBsAg-induced Th1 immune response

Method used

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  • Vaccine for controlling persistent infection of hepatitis B virus
  • Vaccine for controlling persistent infection of hepatitis B virus
  • Vaccine for controlling persistent infection of hepatitis B virus

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Induced expression, detection and identification of recombinant Hansenula cells expressing hepatitis B surface antigen:

[0056] Construct and induce the expression of HBsAg according to the prior art. First, HBsAg codons were optimized, using the codons favored by Hansenula cells. Conventional PCR method was used to amplify the HBsAg fragment, upstream primer 5'-GATCTTTAAA AACAA AATGGAGAACATCACCCTCTG-3, downstream primer: 5'-CGGAATTCCTATTAGATGTACACCC-3'. The optimized gene sequence was synthesized by bridging PCR. The target gene was subcloned into the multi-copy expression vector pDGXHP2.0 by enzyme digestion and ligation to form a 4-copy recombinant expression plasmid. The above-mentioned recombinant expression plasmid and pDGXHP2.0 plasmid negative control were transformed into Hansenula spp. The colony was transferred into 10 ml of MDL (0.14% amino-free yeast nitrogen source, 0.5% ammonium sulfate, 2% glucose) liquid medium for subculturing on a shaker at 33°C. ...

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Abstract

The invention belongs to the biotechnology field, and relates to a vaccine for controlling the persistent infection of hepatitis B virus. According to the invention, a Hansenula polymorpha cell that is heat inactivated and expresses hepatitis B surface antigen is adopted as the hepatitis B vaccine, wherein the HBsAg is an antigen part of the vaccine and the Hansenula polymorpha cell is an adjuvant part of the vaccine. Animal immunity experiment shows that: the vaccine can induce the aggregation of immune cells to the spleen, promote the maturation of DCs, and both induce Th1 immune response and enhance Th2 immune response in mice, including total IgG, IgG1, IgG2a, specific CTL activity, and the production ability of antigen-specific IFN-gama. The invention can make up for the deficiency of the impossibility of inducing Th1 immune response for traditional vaccines that take aluminium hydroxide as an adjuvant, and is helpful to the control of the persistent infection of hepatitis B virus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a novel strategy for Hansenula cells to enhance HBsAg-induced Th1 immune response, in particular to a vaccine for controlling persistent infection of hepatitis B virus. Background technique [0002] Studies have shown that the vaccine is one of the most effective means of controlling the spread of hepatitis B. At present, the hepatitis B vaccine used at home and abroad is mainly composed of hepatitis B surface antigen (HBsAg) and adjuvant. The immunogenicity of hepatitis B surface antigen (HBsAg) alone is weak, and the adjuvant is used to enhance the immunogenicity of the antigen. Currently, the only adjuvant approved for clinical application is aluminum hydroxide (alum), which mainly enhances the Th2 immune response, mainly humoral immunity. [0003] For patients with chronic HBV infection, practice has shown that the body's immune response cannot effectively eliminate pathogens, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/29A61K39/39A61P31/20C12N15/81C12N15/51
Inventor 袁正宏卞广林
Owner FUDAN UNIV
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