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Tumor DNA vaccine taking mucin 1 and survivin as targets and viral vector vaccine

A technology of vaccinia virus and backbone vector, which is applied in the field of tumor DNA vaccine and virus vector vaccine, and can solve the problems of weak immunogenicity, poor immunogenicity and unsatisfactory clinical effect of DNA vector vaccine

Active Publication Date: 2015-01-07
CHANGCHUN BCHT BIOTECH +1
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AI Technical Summary

Problems solved by technology

From the discovery of MUC1 to the present, a lot of vaccine research work has been carried out with it, and some vaccines have entered the human clinical research stage, but most of them are traditional polypeptide, protein or dendritic cell (DC) vaccines, Most of these vaccines have problems such as poor immunogenicity or difficulty in preparation; the genetic vaccines that have entered the clinic are mainly recombinant poxvirus vaccines. Although no toxicity has been detected, there are still relatively large safety hazards in primary vaccination-boosting of recombinant poxvirus vaccines directly. , and the clinical effect is not very satisfactory; DNA carrier vaccine is still basically in the preclinical research stage due to its weak immunogenicity
[0007] The previous studies of the inventors of the present invention have shown that increasing the number of VNTRs can enhance the immune effect of MUC1 to a certain extent (see Zhang S et al., 2008), for example, the immune response caused by 33 copies of the MUC1 VNTR tandem repeat sequence is significantly stronger than that of 2 Therefore, the present invention selects the tandem repeat sequence MUC1VNTR containing 33 as antigen to design a vaccine, and expresses it with S8 to construct a fusion expression vaccine to enhance the broad-spectrum of vaccine immunity. and effectiveness, the combination of survivin and MUC1VNTR for tumor therapy has not been reported

Method used

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  • Tumor DNA vaccine taking mucin 1 and survivin as targets and viral vector vaccine
  • Tumor DNA vaccine taking mucin 1 and survivin as targets and viral vector vaccine
  • Tumor DNA vaccine taking mucin 1 and survivin as targets and viral vector vaccine

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Embodiment Construction

[0032] 1. The preparation method and conditions of the above-mentioned fragment 33M and S8

[0033] Preparation of 1.33M

[0034] MUC1VNTR is a repeat fragment containing 60 bases, GenBank number is NM_002456 (SEQ ID NO: 1). Two pairs of primers were designed according to the base sequence of a VNTR, and a VNTR (1m) repeat region fragment was synthesized by overlapping extension PCR (SOE PCR), and then digested with restriction endonucleases SalI and XhoI to produce the same sticky end characteristics, sequentially connected to construct 33M gene fragments. The specific method is as follows:

[0035] 1) Overlap extension PCR technology

[0036] The PCR reaction conditions are: 95°C for 20s, 55°C for 20s, 72°C for 30s, for 30 cycles, using primers P1 (SEQ ID NO:2) and P2 (SEQ ID NO:3) to amplify one copy of MUC1VNTR without ATG (abbreviated as m), using P2 and P3 (SEQ ID NO: 4) as primers to amplify 1 copy of MUC1VNTR (abbreviated as Am) fragment containing ATG.

[0037] 2...

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Abstract

The invention discloses a tumor deoxyribonucleic acid (DNA) vaccine taking mucin 1 and survivin as targets and a viral vector vaccine. The invention relates to the field of tumor DNA vaccines and viral vector vaccines, in particular to recombinant DNA vector VR-S8 and VR-MS, recombinant adenovirus Ad-S8 and Ad-MS, and recombinant poxvirus MVA-MS vaccines, and application of the optimum combination of immune ways between DNA vaccines and viral vector vaccines and between the viral vector vaccines in preparing antitumor vaccines.

Description

technical field [0001] The invention relates to the field of tumor DNA vaccine and virus vector vaccine. Specifically, the present invention relates to recombinant DNA vector VR-S8 and VR-MS, recombinant adenovirus Ad-S8 and Ad-MS and recombinant poxvirus MVA-MS vaccine, and DNA vaccine and viral vector vaccine and viral vector vaccine The application of the optimized combination of immunization methods in the preparation of anti-tumor vaccines. Background technique [0002] Survivin is a member of the inhibitor of apoptosis protein (IAP) family, has dual functions of anti-apoptosis and regulation of cell division, and is widely expressed in various embryonic tissues and cancer cells, but not in normal terminally differentiated cells . These characteristics make survivin a new target in the fields of tumorigenesis, development and therapy. [0003] Current research shows that survivin has potential value in tumor gene therapy: its tumor-specific expression is positively c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/62C12N15/85C12N7/01A61K39/00A61K48/00A61P35/00C12R1/93
Inventor 孔维张海红于湘晖于永慧
Owner CHANGCHUN BCHT BIOTECH
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