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Tumor vaccine and preparation method thereof

A tumor vaccine and tumor vaccine technology, applied in the field of genetic engineering, can solve problems such as being unsuitable for extensive clinical application, and achieve the effect of overcoming obvious side effects and good curative effect

Inactive Publication Date: 2014-10-29
TIANJIN MEIDE BAITAI MEDICAL TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Autologous lung cancer cell tumor vaccine has autologous specific tumor antigen, which is safer and more effective than allogeneic tumor cell tumor vaccine, but it is not suitable for wide clinical application

Method used

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  • Tumor vaccine and preparation method thereof
  • Tumor vaccine and preparation method thereof

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Experimental program
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Effect test

specific Embodiment 1

[0077] The A549 lung cancer cell line with the highest frequency of class I antigen in the Chinese population was selected as the object. Extraction of GM-CSF-mRNA and synthesis of cDNA, PCR amplification of GM-CSF gene, cloning of GM-CSF cDNA, construction of GM-CSF recombinant Sendai virus vector, preparation of GM-CSF recombinant Sendai virus packaging cells, Infection of human lung cancer cell line A549, 60 The Co treatment instrument performs cell biology, molecular biology, virology and other technologies to irradiate and inactivate tumor cells, and obtains genetically engineered human lung cancer cell tumor vaccines with GM-CSF expression activity. ℃~65℃ warm bath for 0.5h~2h for heat treatment. 60 The Co therapeutic apparatus was irradiated to inactivate, and the irradiated cells were named GM-CSF genetically engineered human lung cancer cell tumor vaccine (A549 / GM-CSF).

specific Embodiment 2

[0078] Different doses of human lung cancer cells modified by granulomatous colony-stimulating factor 60 Co irradiated cells lost their proliferative ability in vitro, and with the increase of culture days, the cell viability decreased and the number of living cells decreased. As shown in Table 2, the survival rate of cells after one week of radiation was 50%, and there was no difference among different radiation doses. The cells irradiated by 60G and 100G all died when cultured to the 25th day. When the genetically engineered human lung cancer cells (HG-1, GM-CSF) irradiated with the above three doses were inoculated into nude mice, no tumor was produced, indicating that the tumorigenicity disappeared.

[0079] Table 1 Gene modification of human lung cancer cells by granulomatous cell colony-stimulating factor 60 Survival rate after CO irradiation

[0080]

[0081]

[0082] Will 60 The granulomatous colony-stimulating factor gene-modified human lung cancer cells tha...

specific Embodiment 3

[0083] The preventive effect of "A549 / GM-CSF tumor vaccine" on mouse tumor models: 6-8 weeks old syngeneic BALB / c mice, the following cells were taken 5×10 5 / Rats were inoculated subcutaneously in the abdomen: ① irradiated and inactivated primary A549 tumor vaccine; ② irradiated and inactivated "A549 / GM-CSF tumor vaccine". Take 1×10 after 7 days 4 The primary A549 tumor vaccine was injected subcutaneously in the other side of the abdomen, and the tumor volume was measured every other day after bearing the tumor. Efficacy of cancer vaccines.

[0084] After 55 days of observation, no tumor growth was seen in the "A549 / GM-CSF tumor vaccine" group, and all survived during the entire observation period (120 days); the mice in other groups could see rapid tumor growth 17 days after the tumor was planted and the mice in the other groups could see rapid tumor growth at 46 days. All died within days.

[0085] The "A549 / GM-CSF tumor vaccine" group was treated with 1×10 4 , 5×10 5...

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Abstract

The invention provides a tumor vaccine and a preparation method thereof. The preparation method comprises: extracting mRNA from human peripheral blood monouclear cells, synthesizing cDNA in vitro, employing polymerase chain reaction technology to clone granulocyte macrophage colony-stimulating factor (GM-CSF) cDNA containing a signal peptide, recombining with sendai virus vector, so as to construct a recombinant sendai virus vector GM-CSF-SV containing granulocyte macrophage colony-stimulating factor cDNA; and employing a packaging cell 293T with relatively high safety to package the recombinant sendai virus vector GM-CSF-SV containing GM-CSF gene, so as to generate he recombinant sendai virus vector which is high in titer and does not have duplication capability; and infecting human lung cancer cell strain A549, so as to establish a gene engineering human lung cancer cell with GM-CSF expression activity, and employing <60>Co to irradiate and inactivate a tumor vaccine and performing heat shock processing, and preparing the GM-CSF gene modified human lung cancer cell tumor vaccine after different irradiation amounts are explored.

Description

technical field [0001] The invention relates to the field of genetic engineering, and more specifically, the invention provides a tumor vaccine and a preparation method thereof. Background technique [0002] Lung cancer is one of the malignant tumors with the highest morbidity and mortality in the world. At present, most of the lung cancer patients in our country are in the middle and late stage, the surgical resection rate is only about 60%, and the 5-year survival rate after operation is only about 50%. As the fourth method of tumor treatment after surgery, radiotherapy and chemotherapy, biological therapy has developed rapidly in recent years. Immunotherapy is a major component of biological therapy. Tumor vaccine is a tumor-specific active immunotherapy. It induces a specific active immune response of the body and enhances the anti-tumor ability of the body, which has been affirmed in animal experiments. Many tumor vaccines have entered clinical trials and have shown g...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00A61P11/00
Inventor 赵钢闫贵芹崔国巍
Owner TIANJIN MEIDE BAITAI MEDICAL TECH DEV CO LTD
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