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DC (dendritic cell) based breast cancer therapeutic vaccine and preparation method thereof

A therapeutic vaccine and breast cancer technology, applied in the fields of bioengineering and biomedicine, can solve the problems of poor tumoricidal ability and weak immune response, and achieve the effects of strong tumoricidal ability, promotion of antigen presentation, and stable traits.

Inactive Publication Date: 2016-06-15
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a DC-based therapeutic vaccine for breast cancer with strong immunogenicity and specificity in view of the defects of weak immune response and poor tumor killing ability in the prior art breast cancer therapeutic vaccine and its preparation method

Method used

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preparation example Construction

[0026] A kind of DC-based breast cancer therapeutic vaccine provided by the present invention, its preparation method comprises the following steps:

[0027] S1. Obtain immature DC;

[0028] S2. Co-cultivating the breast cancer antigen complex and the immature DC in the DC medium for 5-9 days; wherein, the breast cancer antigen complex is obtained by co-culturing the breast cancer cells and the MUC1 monoclonal antibody complex after extinguishing fire;

[0029] S3, then adding tumor necrosis factor-α to continue culturing until the DCs are mature and loaded with breast cancer antigen complexes.

[0030] In step S1, immature DCs can be derived from human blood, but since immature DCs only account for 0.5-1.0% of the total number of monocytes in the blood, the number is very small; therefore, in a preferred embodiment of the present invention, induction of monocytes Obtain immature DC by means of cells, the specific steps are:

[0031] S11, isolating mononuclear cells from per...

Embodiment 1

[0052] The preparation method of the breast cancer therapeutic vaccine based on DC comprises the following steps:

[0053] S1. Acquisition of breast cancer antigen complex:

[0054] S11. MCF-7 breast cancer cells in the logarithmic growth phase were purchased from the Hematology Hospital of the Chinese Academy of Medical Sciences;

[0055] S12. Inactivate the breast cancer cells in step S11 by freezing and thawing;

[0056] Take breast cancer cells in the logarithmic growth phase, wash them twice with normal saline, centrifuge at 1800 rpm, resuspend the cell solution in PBS solution, and obtain 1×10 7 cells / ml of breast cancer cell suspension, take 20 μL of cell suspension and add it to 100 μL of normal saline, then put it into liquid nitrogen, put it in a 37°C water bath after 5 minutes, and put it in again after the breast cancer cells are completely melted. In liquid nitrogen, repeat 3 times, then filter through a microporous membrane, collect the filtrate, and store it a...

Embodiment 2

[0068] The difference from Embodiment 1 provided by the present invention is that in this embodiment,

[0069] The concentration of immature DC was 2 × 10 5 a / ml;

[0070] The concentration of breast cancer cell suspension was 0.5×10 7 a / ml;

[0071] The concentration of MUC1 monoclonal antibody complex was 0.5 μg / (1×10 5 ) cells;

[0072] The concentration of tumor necrosis factor-α is 800U / ml;

[0073] The volume ratio of immature DC to breast cancer antigen complex was 4:1.

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Abstract

The invention relates to a DC (dendritic cell) based breast cancer therapeutic vaccine and a preparation method thereof. The preparation method of the DC based breast cancer therapeutic vaccine comprises steps as follows: immature DCs are acquired; a breast cancer antigen compound and the immature DCs are subjected to co-culture in a DC culture medium for 5-9 days; a tumor necrosis factor-alpha is added for continuous culture until the DCs are mature and are loaded with the breast cancer antigen compound, wherein the breast cancer antigen compound is prepared from inactivated breast cancer cells and a MUC1 monoclonal antibody compound through co-culture. The DC based breast cancer therapeutic vaccine has higher immunogenicity and can be used for regulating the body immune mechanism for breast cancer very well.

Description

technical field [0001] The invention relates to the fields of bioengineering and biomedicine, in particular to a DC-based breast cancer therapeutic vaccine and a preparation method thereof. Background technique [0002] Breast cancer is the most common malignant tumor among women, and its incidence is increasing year by year. The existing breast cancer treatment process has greatly damaged the patient's immune system, resulting in a bottleneck that cannot be broken through in the tumor treatment process, poor tumor killing ability, and the tumor cannot be completely cured. At present, cytokine-induced killer cells are immunocompetent cells with cytotoxicity; dendritic cells are the most potential antigen-presenting cells, and play an important role in initial immunity and acquired immunity; dendritic cells are It is known that the cell with the strongest antigen presentation ability in the body can capture the antigen and transmit the information to T and B lymphocytes, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61K39/00A61P35/00C12N5/0784
CPCA61K39/0011A61K39/395A61K2039/505A61K2039/5154A61K2039/585C12N5/0639C12N2501/25C12N2501/998
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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