Recombined adenovirus vaccine for preventing AIDS

A technology of adenovirus and virus, applied in the field of new vaccines, can solve the problems of low HIV immune protection response and limited immune intensity, and achieve the effect of good safety and high expression level

Inactive Publication Date: 2012-07-11
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limited immune strength and the difficulty of multiple booster immunizations with recombinant vector vaccines, the results of animal models and human trials have shown low immune protection responses to HIV

Method used

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  • Recombined adenovirus vaccine for preventing AIDS
  • Recombined adenovirus vaccine for preventing AIDS
  • Recombined adenovirus vaccine for preventing AIDS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Selection and modification of antigen

[0024] 1. Selection of target gene

[0025] At present, the main HIV-1 epidemic strain in China is B / C recombinant HIV-1.

[0026] The gagpol gene sequence of the selected HIV-1 Chinese epidemic strain is as SEQ ID NO: 6,

[0027] The selected HIV-1 Chinese epidemic strain env gene sequence is as SEQ ID NO: 7,

[0028] The HIV-1 target gene we used to construct the AIDS vaccine here is based on the gene sequence of the above-mentioned B / C recombinant subtype, and then the full sequence of gagpol was artificially synthesized. The amino acid sequence expressed by the synthetic gene is similar to that of the wild one. The gagpol gene expressed by the HIV-1 Chinese epidemic strain has the same amino acid sequence, but the gene expression efficiency is greatly improved.

[0029] The modified gagpol and wild env genes are used as target antigen genes, and their expression products constitute the most important structural protein of HIV-1, so they...

Embodiment 2

[0104] Construction of recombinant adenoid vaccine A-GPE

[0105] 1. Construction process

[0106] The gagpol gene used to construct the recombinant adenoid vaccine is the modified gagpol gene in Example 1, the env gene used to construct the recombinant adenoid vaccine is the wild-type env gene, and the adenovirus is a replication-deficient adenovirus in humans.

[0107] The gagpol and env genes of the aforementioned HIV-1 Chinese epidemic strain (regional) were cloned into the intermediate plasmid pDC316 to form pDC316-GPE. Such as figure 1 Shown: In the pDC316-GPE plasmid, the modified gagpol gene and wild-type env gene were ligated into the shuttle plasmid pDC316 using the polyclonal restriction site in the shuttle plasmid to obtain the plasmid pDC316-GPE.

[0108] The full sequence of the recombinant shuttle plasmid pDC316-GPE is as SEQ ID NO:1

[0109] Using Lipofection2000 (product of INVITROGEN), pDC316-GPE and adenovirus packaging plasmid were co-transfected into HEK293 cells. ...

Embodiment 3

[0166] Study on the antigenicity of A-GPE

[0167] Test materials and equipment, test methods and steps: the same as in Example 2 for the expression of antigen in mammalian cells by recombinant adenoids.

[0168] Results: In order to detect whether the antigenicity of A-GPE is consistent with that of the epidemic strains in China, we infected HEK293 cells with A-GPE to express HIV-1 antigen, and used Western blotting to confirm whether the expressed antigen was The antiserum of the epidemic strain in China (from the HIV-1 high-incidence area of ​​Guangxi Zhuang Autonomous Region) was identified in the Western blot test. At the same time, the virus antigen of the Chinese epidemic strain of HIV-1 (from the HIV-1 high-incidence area of ​​Guangxi Zhuang Autonomous Region) was used as a positive control. The test results are as Figure 4 Shown.

[0169] The protein expressed by A-GPE can be recognized by China's epidemic (regional) antisera and has the same antigenicity as the correspon...

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Abstract

The invention provides a replication defective recombinant adenovirus vector vaccine for preventing AIDS. The recombinant adenovirus vaccine contains a nucleotide sequence which can encode human immunoddficiency virus-1 (HIV-1) structural proteins. A transcription unit contained in the recombinant adenovirus vaccine encodes HIV-1B / C recombinant complete core protein Gag, enzyme proteins coding Pol and outer membrane protein Env. In addition, in order to improve the efficiency of gene expression, the recombinant adenovirus ultimately selects modified Gag and Pol genes and wild type Env gene.

Description

Technical field [0001] The present invention relates to a new vaccine, in particular, the vaccine is a recombinant gland containing artificially modified Gag, Pol and wild Env nucleotide sequences encoding human immunodeficiency virus-1 (HIV-1) structural protein virus. These vaccines are used as transcription units for the biosynthesis of viral protein antigens when administered in vivo. Background technique [0002] AIDS is considered to be the world’s largest infectious disease that directly threatens human health. The "2004 Annual Report on the Global AIDS Epidemic" published by the United Nations Joint Programme on HIV / AIDS pointed out that there are currently 38 million people living with HIV in the world, and a total of 2.9 million in 2003 People died of AIDS. In 2003, 1.1 million people in Asia were infected and more than 500,000 died from AIDS. There are currently 7.4 million people living with HIV in the region. At the end of 2003, there were about 840,000 people liv...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/12A61K48/00A61P31/18
Inventor 于湘晖孔维詹杨于彬
Owner JILIN UNIV
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