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Recombinant BCG vaccine based on human MUC1 repetitive sequence and GM-CSF fusion expression

A GM-CSF, repeating sequence technology, applied in the field of recombinant BCG vaccine, can solve the problems of insufficient immune response and protection, and achieve the effect of reducing tumor formation rate, significant immune preventive effect, and improving immunogenicity

Inactive Publication Date: 2012-09-26
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0031] Chung et al. used BCG as an expression carrier of foreign genes for anti-tumor research, immunized hu-PBL-SCID mice with full-length MUC1 and human interleukin 2 (IL-2) recombinant BCG vaccine, and vaccinated once every 2 weeks for a total of Three times, it was found that it can inhibit the growth of breast cancer, but the immune response and protection it induces are still insufficient, which may be related to factors such as the full-length sequence of MUC1 affecting its immune effect

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  • Recombinant BCG vaccine based on human MUC1 repetitive sequence and GM-CSF fusion expression
  • Recombinant BCG vaccine based on human MUC1 repetitive sequence and GM-CSF fusion expression
  • Recombinant BCG vaccine based on human MUC1 repetitive sequence and GM-CSF fusion expression

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Embodiment Construction

[0063] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments, which are explanations rather than limitations of the present invention.

[0064] The present invention utilizes gene recombination technology to obtain a new expression vector MUC1VNTRs-GM-CSF by cloning the continuous repeat sequence of the human MUC1 concatemer and fusing it with the GM-CSF gene. The vector expresses the fusion protein MVNTRs-GM-CSF to Improving the immunogenicity of MUC1; the human MUC1 tandem continuous repeat sequence described in the present invention is a multi-tandem continuous repeat sequence, specifically taking 1 tandem, 4 tandem and 8 tandem as examples, which are respectively expressed as MUC1VNTR1-GM -CSF, MUC1VNTR4-GM-CSF, MUC1VNTR8-GM-CSF, for the convenience of description, the above three concatenations are described as MUC1VNTR1 / 4 / 8-GM-CSF;

[0065] The above-mentioned fusion gene is transfected into...

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Abstract

The invention discloses a recombinant BCG vaccine based on human MUC1 repetitive sequence and GM-CSF fusion expression. Firstly, MUC1 repetitive sequence multimers and fusion genes connected with GM-CSF genes by connecting DNA are constructed, a signal peptide sequence and a Kozak sequence are further connected, and an expression vector is constructed; and the constructed fusion genes are transfected to BCG vaccines, and applied to the preparation of anti-tumor vaccine or anti-tumor medicament by using MUC1 as a target spot. The recombinant BCG vaccine realizes effective combination of antibody, adjuvant and vaccine vector so that the immunogenic property of the MUC1 tumor vaccine achieves more effective and durable effect; and simultaneously the recombinant BCG vaccine is safe, has no toxic and side effects, and can induce specific T cell immune response and durable secretion expression of the human MUC1 repetitive sequence and GM-CSF fusion protein.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and relates to the preparation of anti-tumor vaccines, in particular to a recombinant BCG vaccine based on the fusion expression of human MUC1 repeat sequence and GM-CSF, and its preparation method and application. Background technique [0002] 1. Research on MUC1 molecules and MUC1 tumor vaccines [0003] 1.1 Molecular biology of MUC1 [0004] Mucin1 mucin (MUC1), also known as polymorphic epithelial mucin (polymorphic epithelial mucin, PEM), DF3 and so on. MUC1 is a highly glycosylated type I transmembrane protein. Under normal circumstances, it is mainly expressed on the glandular surface of epithelial cells in various tissues and organs, with apical expression and polar distribution; MUC1 is abnormally expressed on the surface of cancer cells. A non-polar distribution. [0005] The MUC1 gene is located at band 21 of the long arm of chromosome 1 and consists of 7 exons with a length ran...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62A61K39/00A61K48/00A61P35/00A61P15/14
Inventor 袁时芳师长宏韩苇张英起凌瑞李南林王廷
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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