AIDS vaccine based on replicative vaccinia virus vector

A vaccinia virus, AIDS technology, applied in the direction of viruses/phages, viruses, viral peptides, etc., can solve the problems of short-term, limited immune system stimulation, and inability to produce infectious viruses.

Inactive Publication Date: 2007-08-22
NAT CENT FOR AIDSSTD CONTROL & PREVENTION CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the non-replicating vector vaccine infects the host cell, it can only carry out a cycle of antigen expression, processing and presentation, and cannot produce infectious virus and start a new round of infection, so its stimulation of the immune system is limited and Short-lived

Method used

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  • AIDS vaccine based on replicative vaccinia virus vector
  • AIDS vaccine based on replicative vaccinia virus vector
  • AIDS vaccine based on replicative vaccinia virus vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Construction of vaccinia virus universal transfer vector pVTT 1.0

[0047] 1. Construction of recombinant plasmid pSC-neo

[0048] Plasmid pIRESneo (purchased from Clontech Company) was first digested with XhoI, then digested with SmaI (reaction temperature: 25°C), filled in with Klenow enzyme, and the 1.2kb target fragment neo-polyA was recovered; the transitional vector plasmid pSC65 (preservation number: CGMCC No.1097) was digested with BglII, blunted with Klenow enzyme, treated with dephosphorylase (CIAP), and the vector was recovered; the two were ligated at 16°C for 4 hours, and transformed into E. coli TOP10. Multiple single colonies were picked, a small amount of plasmid was extracted, identified by XbaI and PstI, and the correct recombinant clone was named pSC-neo.

[0049] 2. Artificially synthesized vaccinia virus vector early promoter PE6 and lacZ fusion fragment.

[0050] Genes were synthesized by overlapping PCR (Overlapping PCR). First, the ...

Embodiment 2

[0054] Embodiment 2: Construction of transfer plasmid pVTT-gagpolenv

[0055] The construction of plasmid pVTT-gagpolenv was carried out in three steps:

[0056](1) Construction of plasmid pVTT-gagpol

[0057] Plasmid pT-gagpol (CGMCC, General Microbiology Center, China Committee for Culture Collection of Microbial Cultures, deposit number CGMCC No.1438) was constructed by the Chinese Center for Disease Control and Prevention of STD and AIDS, and contained the gagpolΔ gene. The plasmid was digested with EcoRI and XmnI, and Klenow The target fragment of 2.9kb was recovered by enzyme filling; the vector pVTT 1.0 was digested with SmaI, dephosphorylated by CIAP, and the linearized vector was recovered. The vector and the fragment were ligated and transformed into Escherichia coli DH5α, and a single colony was picked to extract the plasmid for enzyme digestion identification, and the correct clone was named pVTT-gagpol. The plasmid pVTT-gagpol was digested with PstI, XbaI, and N...

Embodiment 3

[0062] Embodiment 3: Screening of recombinant virus

[0063] The tkL and tkR regions of the transfer plasmid pVTT-gagpolenv undergo homologous recombination with the vaccinia virus Tiantan strain, so that the target genes gagpolΔ and gp140TM and marker genes neo and lacZ are recombined into the TK sequence of the vaccinia virus genome. In the presence of G418, due to the absence of homologous recombination in the selective pressure molecule, the marker genes neo and lacZ temporarily remain in the genome, and the low-melting point agarose with X-gal and neutral red can be used to pick out Blue recombinant vaccinia virus containing both the gene of interest and the selectable marker (viruses that have not undergone recombination are inhibited from growth due to the presence of G418). Then screened under the condition of no G418, the blue recombinant vaccinia virus itself will lose the neo gene and The recombinant vaccinia virus containing only the target gene was obtained by us...

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Abstract

The present invention relates to replicative live AIDS carrier vaccine expressing HIV antigen and its use. The vaccine is constructed based on replicative vaccinia virus, such as vaccinia virus Tiantan strain. The replicative live AIDS carrier vaccine can induce high level HIV resisting body fluid and cellular immune response. The present invention provides the carrier for constructing the AIDS vaccine, and also relates to immunizing process with the AIDS vaccine.

Description

technical field [0001] The invention relates to the field of antiviral immunology. More specifically, the present invention relates to a vaccine against Human Immunodeficiency Virus (Human Immunodeficiency Virus, HIV) based on replicating vaccinia virus vector and its preparation method and use. The AIDS vaccine of the invention can induce high-level anti-HIV humoral and cellular immune responses. Background technique [0002] Acquired Immunodeficiency Syndrome (AIDS) is abbreviated as AIDS, which is an infectious disease caused by Human Immunodeficiency Virus (Human Immunodeficiency Virus, HIV) infection. Since the first case was discovered in 1981, AIDS has been spreading around the world at an alarming rate and has become one of the most serious viral diseases that endanger human life and health in the world. The AIDS epidemic has brought a huge impact on the social and economic development of the world. In some developing countries, HIV infection has shortened life ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/125A61K48/00C12N15/89C12N15/49A61P31/18
CPCC12N2740/16234C12N2710/24143C12N2740/16122A61K39/21A61K2039/545A61K2039/54A61K2039/5256C12N2740/16222C12N2740/16134C07K14/005A61K2039/575A61K39/12A61P31/18
Inventor 邵一鸣刘颖刘建元
Owner NAT CENT FOR AIDSSTD CONTROL & PREVENTION CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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