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Transgenic natural killer cell and preparation method and application thereof

A natural killer cell and genetically modified technology, applied in the field of biopharmaceuticals, can solve the problems of tumor cell escape, inhibition of killing effect, poor curative effect, etc., and achieve the effects of avoiding immune escape, enhancing killing activity, and high infection rate

Active Publication Date: 2017-02-22
SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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Problems solved by technology

[0007] The purpose of the embodiments of the present invention is to overcome the above-mentioned deficiencies in the prior art, and provide a transgenic natural killer cell and a preparation method thereof to solve the problem that the current transgenic natural killer cells escape from tumor cells due to a tumor escape mechanism, and their killing effect is limited. Suppressed technical issues
[0008] Another object of the embodiments of the present invention is to provide a tumor drug preparation to solve the technical problem of poor curative effect of the existing biological therapy tumor drug preparation using NK cells

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  • Transgenic natural killer cell and preparation method and application thereof
  • Transgenic natural killer cell and preparation method and application thereof
  • Transgenic natural killer cell and preparation method and application thereof

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preparation example Construction

[0033] On the other hand, on the basis of the transgenic natural killer cells of the embodiments of the present invention described above, the embodiments of the present invention also provide a preparation method of the transgenic natural killer cells of the embodiments of the present invention. The preparation method of the embodiment of the present invention comprises the following steps:

[0034] NK cells were simultaneously infected with the first lentivirus and the second lentivirus.

[0035] Wherein, the first lentivirus contains the above-mentioned chimeric antigen receptor gene targeting mesothelin antigen. In one embodiment, the first lentivirus is prepared according to the following method:

[0036] S11: Mix the Meso / scFv-CD8-CD28-CD3ζ gene fragment with the entry vector, so that the Meso / scFv-CD8-CD28-CD3ζ gene fragment is embedded in the entry vector to obtain the Meso / scFv-CD8-CD8- The entry vector of CD28-CD3ζ gene fragment;

[0037] S12: mixing and recombini...

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Abstract

The invention provides a transgenic natural killer cell and a preparation method and application thereof. The provided transgenic natural killer cell comprises a targeting mesothelin antigen-specific chimeric antigen receptor gene and a tyrosine phosphatase SHP gene in a silencing NK cell. The preparation method includes the step of infecting the NK cell with slow viruses containing the targeting mesothelin antigen-specific chimeric antigen receptor gene and slow viruses containing the tyrosine phosphatase SHP gene in the silencing NK cell simultaneously. By the combined action of the targeting mesothelin antigen-specific chimeric antigen receptor gene and the tyrosine phosphatase SHP gene in the silencing NK cell, the transgenic natural killer cell is capable of targeting and identifying tumor cells in a specific and targeting mode to avoid the immune escape of the tumor, and the activity on killing the tumor cells is enhanced.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to a transgenic natural killer cell, a production method thereof and a pharmaceutical preparation containing the transgenic natural killer cell. Background technique [0002] Natural killer cells (NK cells) are important effector cells in innate immunity and play an important role in preventing viral infection and inhibiting cancer development. NK cells have a strong killing effect on tumor cells and are non-MHC-restricted, and their recognition of tumor cells mainly depends on the mutual cross-regulation of their surface activating receptors and inhibitory receptors. [0003] Normal cells send inhibitory signals to NK cells to control their killing function due to the combination of their own MHC-I molecules on their surface and the inhibitory receptors on the surface of NK cells. Downregulation of MHC-1 in tumor cells sensitizes them to the cytotoxic effect...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867A61K35/17A61P35/00
CPCA61K35/17C07K14/7051C07K14/70517C07K14/70521C07K16/28C07K16/3015C07K2317/622C07K2319/00C12N5/0646C12N9/16C12N15/86C12N2740/15043C12Y301/03048
Inventor 不公告发明人
Owner SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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