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Preparation method of prostatic cancer specific DC cell vaccine which is insensitive to TGF-belta

A prostate cancer, TGF-technology, applied in the field of DC cell vaccine preparation, can solve the problems of prostate cancer loss and treatment failure, and achieve significant clinical significance and application value

Inactive Publication Date: 2010-03-31
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Claims
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Problems solved by technology

At present, the clinical treatment of prostate cancer is mainly based on anti-androgen therapy, but because prostate cancer is a heterogeneous tumor, as the disease progresses, the dependence of prostate cancer on androgen will change and be lost, eventually leading to treatment failure
So far, there is no effective treatment, so finding an effective and specific treatment for prostate cancer is an urgent problem to be solved in the clinical treatment of urology.

Method used

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  • Preparation method of prostatic cancer specific DC cell vaccine which is insensitive to TGF-belta
  • Preparation method of prostatic cancer specific DC cell vaccine which is insensitive to TGF-belta
  • Preparation method of prostatic cancer specific DC cell vaccine which is insensitive to TGF-belta

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Experimental program
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Embodiment

[0033]1) Construction of a retrovirus containing a dominant negative TGF-β type II receptor (TβRIIDN) plasmid: transfect 293 packaging cells with a retrovirus containing a TβRIIDN plasmid, and co-transfect for 12 hours at 32°C in 10% DMEM medium Incubate overnight at 37°C, rinse with PBS and retransfect 293 cells under the same conditions for 24 hours, collect the supernatant, and obtain recombinant retroviruses containing TβRIIDN.

[0034] 2) Isolation, culture and identification of DC cells loaded with prostate cancer antigen: DC cells were obtained from the fresh bone marrow of C57BL / 6 mice by red blood cell depletion, and added with rhIL-4 (1000U / ml) and GM-CSF (1000U / ml) half of the medium was changed every other day, and the non-adherent cells were harvested by gradient centrifugation on the 6th day as immature DC cells. Repeated freezing and thawing obtained TRAMP-C2 cell lysates, and the lysates repeatedly stimulated immature DC cells to obtain mature DC cells loaded w...

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Abstract

The invention relates to preparation method of prostatic cancer specific DC cell vaccine which is insensitive to TGF-belta, which can treat prostatic cancer effectively, and is prostatic cancer specific immunization gene treatment with genuine with truly implemented combination of active immunity and adoptive cellular immunity. The technical scheme of the present invention applied is: 1) constructing retrovirus containing dominant negative TGF-belta II type (T belta RIIDN) plasmid; 2) separating, culturing and identifying DC cells loaded with prostatic cancer antigen; 3) transfection of DC cells to obtain T belta RIIDN-DC cells; 4) vitro experiments of the T belta RIIDN-DC cells; 5) antitumor estimate of the DC vaccine in vivo; 6) CTL activity detection in vivo.

Description

1. Technical field: [0001] The invention relates to the preparation of a tumor DC vaccine, in particular to a preparation method of a TGF-beta insensitive prostate cancer-specific DC cell vaccine. 2. Background technology: [0002] Prostate cancer is the most common malignant tumor in European and American countries, and its mortality rate is second only to lung cancer. With the development and progress of our society and the extension of the average life expectancy of the population, the incidence of prostate cancer has shown a significant increase trend. At present, the growth rate of the incidence of prostate cancer in my country has ranked first in the growth rate of malignant tumors. Prostate cancer is characterized by a high incidence of latent cancer. More than 75% of prostate cancer patients have metastasized when clinical symptoms appear, which is not suitable for surgical treatment, and the effects of chemotherapy and radiotherapy are not satisfactory. At present...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00A61P13/08C12N7/01C12N5/07C12N5/10
Inventor 王福利秦卫军袁建林田丰温伟红宋朝军董青川王龙信郭应禄王禾
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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