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Nucleic acid vaccine for preventing AIDS

A technology of nucleic acid sequence and nucleotide sequence, which is applied in the field of new vaccines to achieve the effects of reducing production costs, expanding production, and improving immune protection

Inactive Publication Date: 2005-06-29
长春百克药业有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

DNA vaccines induce both humoral and cellular immunity like live attenuated vaccines, but do not have the potential risk of the latter

Method used

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  • Nucleic acid vaccine for preventing AIDS
  • Nucleic acid vaccine for preventing AIDS
  • Nucleic acid vaccine for preventing AIDS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The selection and modification of embodiment 1 antigen

[0017] 1. Selection of the target gene

[0018] At present, the main HIV-1 epidemic strain in China is B / C recombinant HIV-1.

[0019] The gagpol gene sequence of the selected HIV-1 epidemic strain in China is shown as SEQ ID NO: 7

[0020] The env gene sequence of the selected HIV-1 epidemic strain in China is shown as SEQ ID NO: 8

[0021] The HIV-1 target gene we use to construct the AIDS vaccine here is based on the gene sequence of the above-mentioned B / C recombinant subtype, and then artificially synthesized the full sequence. The amino acid sequence expressed by the synthesized gene is the same as that of HIV-1 China The amino acid sequences expressed by the gagpol and env genes of the epidemic strains are consistent, but the gene expression efficiency is greatly improved.

[0022] With the above-mentioned gagpol and env genes as the genes of target antigens, their expression products constitute the most...

Embodiment 2

[0138] Example 2 Construction of Nucleic Acid Vaccine

[0139] 1. Selection of expression vector

[0140] 1.1 Overview.

[0141] The construction of the vector (VR) of the DNA vaccine D-GPEi draws on the multiple cloning sites (including XbaI and BamHI, etc.) Factors, Intron A and BGH PolyA translation termination signal etc. conventional components. VR1012 is a carrier officially approved by the US FDA and can be used in clinical trials of human gene vaccines. The completed clinical trials have shown that its application in humans is safe.

[0142] The construction idea of ​​this DNA vaccine is: use VR1012 as a template, copy the CMV promoter, IntronA and BGH polyA signal, and then clone the expression frame back into VR1012 by using the appropriate restriction site. Therefore, the new plasmid is composed of two CMV promoters, one kanamycin resistance gene, one prokaryotic cell high copy factor, two IntronA and two BGH polyA signals and other components. All parts of the ...

Embodiment 3

[0240] Example 3 Antigenicity Study of Plasmid D-GPEi

[0241] Objective: To investigate the antigenicity of plasmid D-GPEi through antigen expression of plasmid D-GPEi in mammalian cells.

[0242] The specific experimental methods and instrumental reagents are the same as those in Example 2 for the antigen expression of the 4 plasmid D-GPEi in mammalian cells.

[0243] Results and Discussion: In order to ensure the antigenicity of this AIDS vaccine is consistent with the antigenicity of HIV-1 epidemic strains in China (regional), we confirmed the gene sequence of HIV-1 epidemic strains in China according to the results of a large number of epidemiological studies. In order to increase the expression efficiency of the antigen in vivo, we modified the gene sequence of the wild-type HIV-1 and designed the gene sequence of the antigen part of the AIDS vaccine, but kept the amino acid residue sequence of the antigen protein unchanged.

[0244] In order to further confirm that the...

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Abstract

The invention relates to a DNA vaccine for preventing AIDS, the vaccine is the recombinant nucleotide of Gag, Pol and Env nucleotide sequence through the artificial modification of structural protein containing encoded Human Immunodificidncy Virus-1 (HIV-1), these vaccines are used as the transcript units for the biological synthesis of the virus protein antigen when applied in vivo. The beneficial effects of the invention include, (1) improved DNA vaccine manufacturing technique, (2) no integration indication detected, (3) the vaccine target antigen includes almost all the HIV-1 structural protein, such as GagPol and Env.

Description

technical field [0001] The invention relates to a new vaccine, especially, the vaccine is a recombinant nucleic acid containing artificially modified Gag, Pol and Env nucleotide sequences encoding human immunodeficiency virus-1 (HIV-1) structural protein. These vaccines serve as transcriptional 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. According to the "2004 Global AIDS Epidemic Annual Report" issued by the United Nations Joint Program on AIDS, there are currently 38 million HIV carriers in the world, and there were 2.9 million in 2003. People die from AIDS. In 2003, a total of 1.1 million people in Asia were infected and more than 500,000 people died of AIDS. Currently, there are 7.4 million HIV carriers in the region. At the end of 2003, there were about 840,000 HIV-infected people in China, including ab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K48/00A61P31/18C12N15/49
Inventor 孔维于晓方田春娟刘滨东
Owner 长春百克药业有限责任公司
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