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Preparation method of tumor immune antigen, and products and application of preparation method

一种免疫抗原、肿瘤的技术，应用在医学工程领域，能够解决肿瘤治疗效果不佳等问题

Inactive Publication Date: 2016-12-14
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is this tumor-induced immune tolerance that leads to poor efficacy of tumor therapy

Method used

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  • Preparation method of tumor immune antigen, and products and application of preparation method
  • Preparation method of tumor immune antigen, and products and application of preparation method
  • Preparation method of tumor immune antigen, and products and application of preparation method

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0096] The present invention also provides a method for tumor immune antigen (especially HSP70), the method is based on the sequential cooling-heating treatment of the present invention, after cold-heating the tumor cells cultured in vitro, separate and purify the tumor cell culture medium tumor immune antigens. Preferably, the preparation method of the present invention comprises:

[0097] A) cooling the cultured tumor tissue and / or tumor cells to obtain cooled tumor tissue and / or tumor cells; wherein, the cooling includes cooling the tumor tissue and / or tumor cells to T1, and -50°C≤T1≤0°C, preferably, -30°C≤T1≤-10°C, more preferably, -25°C≤T1≤-15°C; most preferably, -20°C≤T1≤-18°C ;

[0098] B) heating the cooled tumor tissue and / or tumor cells obtained in A), so as to obtain tumor tissue and / or tumor cell cultures containing tumor immune antigen groups; wherein, the heating includes obtaining in A) The cooled tumor tissue and / or tumor cells are heated to T2, and 37°C

Embodiment 1

[0133] Embodiment 1 cold-heat treatment improves the survival rate of mice

[0134] The mice in the cold-heat treatment group (n=16) were treated with cold-heat treatment, wherein the temperature was lowered to -20°C within 10 minutes, the temperature was lowered for 5 minutes, and the temperature was heated to 50°C within 10 minutes for 20 minutes.

[0135] The mice in the blank control group (n=16) did not receive treatment.

[0136] Observe the living conditions of the cold-heat treatment group and the blank control mice for one year ( figure 1 ). Sixteen of the cold-heat treated mice survived longer than 66 days after inoculation, and finally 11 mice survived to one year. However, the survival time of the 16 mice in the control group did not exceed 64 days after inoculation.

Embodiment 2

[0137] Example 2 Cold-Heat Therapy Promotes the Release of HSP70 from Local Tumors

[0138] After 1 day of cold-heat treatment and single heat treatment in Example 3, and 22 days after inoculation, orthotopic tumor sections were taken for immunohistochemistry. ( figure 2 ) Brown (light color) is the specific staining of HSP70 protein by the antibody, and blue (dark color) is the staining of the cell nucleus by hematoxylin.

[0139] In the control group, it was seen that some isolated HSP70 existed, most of which existed in the cells and were not released. This kind of HSP70 mainly played a role in promoting the survival of tumor cells.

[0140] Regional release of HSP70 could be observed in the single heat treatment group and combined cold and heat treatment group, but in the single heat group, the release area of ​​HSP70 was relatively limited, and some areas with intact nuclei did not release HSP70;

[0141] In the cold-heat treatment group, necrotic death and cell fragme...

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Abstract

The invention discloses a preparation method of tumor immune antigen, and products and application of the preparation method. Concretely, the inventor finds that after local sequential cooling-heating treatment (or cold-heat treatment) is carried out on tumor tissues and / or cells, the tumor tissues and / or cells release a great deal of tumor immune antigen such as heat shock protein 70 both in vivo and in vitro, and the release amount is significantly higher than the amount released by the tumor tissues and / or cells which are only subjected to pure thermal stimulus. After the sequential cooling-heating treatment is carried out, the obtained tumor immune antigen is capable of effectively activating the tumor immune system in a body, such as immunosuppressive cells can be effectively converted into mature dendritic cells, so that the presentation of the immune antigen is improved and tumor immunity is activated.

Description

technical field [0001] The invention belongs to the field of medical engineering. Specifically, the invention relates to a thermophysical method for stimulating tumor immune antigens, especially a method for in vivo heat shock protein 70 (HSP70) levels in tumor tissues. Background technique [0002] Malignant tumor cells form a stable tumor immune tolerance pattern by reducing their own immunogenicity, down-regulating the expression of co-stimulatory molecules on the cell surface, releasing a large number of immunosuppressive factors, and recruiting a variety of immunosuppressive cells to protect tumor cells from escaping the body's immune surveillance and promote tumor development. It is this tumor-induced immune tolerance that leads to poor efficacy of tumor therapy. At the same time, most malignant tumors will cause different degrees of immunosuppression, in order to facilitate the tumor to escape the body's immune surveillance and attack, and promote tumor progression. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12M1/38C12M1/36C12M1/34
CPCC07K14/47C12M41/00C12M41/12C12M35/00C12M41/48
Inventor 徐学敏刘苹张爱丽白景峰孙建奇邹金成
Owner SHANGHAI JIAOTONG UNIV
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