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Preparation method of tumor vaccine for removing regulatory T cells

A tumor vaccine and regulatory technology, which is applied in the direction of anti-tumor drugs, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve the problems of tumor vaccine effect decline, achieve improved tumor killing effect, high removal efficiency, and tumor killing good effect

Inactive Publication Date: 2019-08-30
朗姿赛尔生物科技(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem solved by the present invention is the problem that regulatory T cells lead to a decrease in the effect of tumor vaccines, and a method for preparing a tumor vaccine that removes regulatory T cells is provided

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for preparing a tumor vaccine that removes regulatory T cells, comprising:

[0030] S1. Collect 90ml of autologous peripheral blood from the patient, use Ficoll-Paque Plus reagent to separate the mononuclear cells therein, and place them in a complete nutrient medium for culture, which is added with penicillin / streptomycin and 10% FBS 1640 medium, and place the medium in CO 2After placing in the cell incubator for 3 hours, collect the suspension, centrifuge to obtain T lymphocyte population, add DC medium to the adherent DC cells and continue to culture, start on the 8th day, collect the culture supernatant containing non-adherent cells, and replace the medium , for a total of two days, the collected supernatants were combined and centrifuged to obtain DC cells;

[0031] S2. DC cells are stimulated with corresponding tumor antigens to sensitize the DC cells in vitro to obtain sensitized DC cells; S3. Add modified nano-tungsten oxide particles to the T lymphocy...

Embodiment 2

[0039] A method for preparing a tumor vaccine that removes regulatory T cells, comprising:

[0040] S1. Collect 80ml of autologous peripheral blood from the patient, use Ficoll-Paque Plus reagent to separate the mononuclear cells therein, and place them in a complete nutrient medium for culture, which is added with penicillin / streptomycin and 10% FBS 1640 medium, and place the medium in CO 2 Place the suspension in the cell incubator for 2 hours, collect the suspension, centrifuge to obtain T lymphocyte populations, and continue to culture the adherent DC cells with DC medium. From the 8th day, collect the culture supernatant containing non-adherent cells, and replace the medium , for a total of two days, the collected supernatants were combined and centrifuged to obtain DC cells;

[0041] S2. DC cells are stimulated with corresponding tumor antigens to sensitize the DC cells in vitro to obtain sensitized DC cells; S3. Add modified nano-tungsten oxide particles to the T lymph...

Embodiment 3

[0048] A method for preparing a tumor vaccine that removes regulatory T cells, comprising:

[0049] S1. Collect 100ml of autologous peripheral blood from the patient, use Ficoll-Paque Plus reagent to separate the mononuclear cells therein, and place them in a complete nutrient medium for culture, which is added with penicillin / streptomycin and 10% FBS 1640 medium, and place the medium in CO 2 After placing in the cell incubator for 5 hours, collect the suspension, centrifuge to obtain T lymphocyte population, add DC medium to the adherent DC cells and continue to culture, start on the 8th day, collect the culture supernatant containing non-adherent cells, and replace the medium , for a total of two days, the collected supernatants were combined and centrifuged to obtain DC cells;

[0050] S2. DC cells are stimulated with corresponding tumor antigens to sensitize the DC cells in vitro to obtain sensitized DC cells; S3. Add modified nano-tungsten oxide particles to the T lympho...

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PUM

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Abstract

The invention relates to the field of tumor vaccine production, and provides a preparation method of a tumor vaccine for removing regulatory T cells. The method is used for solving the problem that the tumor vaccine effect is reduced due to the regulatory T cells. The preparation method of the tumor vaccine for removing regulatory T cells comprises the following steps: adding modified nano tungsten oxide particles into a T lymphocyte population, wherein the weight ratio of the nano tungsten oxide particles to the T lymphocyte population is 1:(100-500), and performing irradiation for 3-10min under visible light; magnetically sorting the irradiated T lymphocyte population to obtain CD8+T lymphocytes; and mixing and culturing sensitized DC cells with the sorted CD8+T lymphocytes, wherein theDC cells carrying tumor antigens present antigen information to the CD8+T lymphocytes and activate the antigen information so as to obtain the specific tumor vaccine. After the T lymphocyte populationis screened out, the regulatory T cells are fully removed by adding the nano tungsten oxide particles and performing magnetic sorting so as to improve the tumor killing effect of the tumor vaccine.

Description

technical field [0001] The invention relates to the field of tumor vaccine production, in particular to a method for preparing a tumor vaccine that removes regulatory T cells. Background technique [0002] Tumor is a malignant disease caused by the imbalance of the two "yin and yang" mechanisms of various carcinogenic factors that induce malignant transformation of cells and the anti-tumor monitoring and killing functions of various immune cells. Traditional tumor treatment mainly achieves the goal of early eradication of tumors and reduction of tumor burden through surgery, chemotherapy and radiotherapy, which has the risk of tumor metastasis and recurrence, and has serious side effects such as damage to normal tissues and low immune function. The chances of tumor metastasis, recurrence and death are extremely high. Tumor immune cell therapy is to restore and enhance the tumor patient's own immune monitoring and tumor killing function, which can effectively kill the remain...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00
CPCA61K39/0011A61P35/00
Inventor 陈忠平
Owner 朗姿赛尔生物科技(广州)有限公司
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