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A specific chimeric antigen receptor gene-modified natural killer cell and its preparation method and application

A chimeric antigen receptor and natural killer cell technology, applied in the field of natural killer cells and their preparation, can solve the problem of insufficient activation of downstream signals of transgenic natural killer cells

Active Publication Date: 2020-07-17
SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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Problems solved by technology

[0006] The main purpose of the present invention is to propose a specific chimeric antigen receptor gene-modified natural killer cell, aiming to solve the problem that the existing transgenic natural killer cell activates downstream signals due to the existence of tumor escape mechanism Insufficient technical issues

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  • A specific chimeric antigen receptor gene-modified natural killer cell and its preparation method and application
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  • A specific chimeric antigen receptor gene-modified natural killer cell and its preparation method and application

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preparation example Construction

[0057] The present invention also provides a method for preparing gene-modified natural killer cells with specific chimeric antigen receptors for preparing the above-mentioned gene-modified natural killer cells with specific chimeric antigen receptors. Wherein, the preparation method includes the following steps:

[0058] Simultaneously infect NK cells with the first lentivirus and the second lentivirus, wherein the first lentivirus contains the NY-ESO-1 specific chimeric antigen receptor gene, and the second lentivirus contains the truncated spleen tyrosine Kinase Syk gene.

[0059] In an embodiment, the first lentivirus is obtained as follows:

[0060] S11: Mix the NY-ESO-1 / scFv-CD8-41BB-DAP12 gene fragment with the entry vector, so that the NY-ESO-1 / scFv-CD8-41BB-DAP12 gene fragment is inserted into the entry vector to obtain Entry vector with NY-ESO-1 / scFv-CD8-41BB-DAP12 gene fragment.

[0061] S12: Mix and recombine the entry vector with NY-ESO-1 / scFv-CD8-41BB-DAP12 gene fragme...

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Abstract

The invention discloses a natural killer cell modified by specific chimeric antigen receptor gene and a preparation method and use thereof. The natural killer cell modified by specific chimeric antigen receptor gene comprises a NY-ESCO-1 specific chimeric antigen receptor gene and a truncated spleen tyrosine kinase Syk gene. The preparation method simultaneously infects the natural killer cell with a slow virus containing the NY-ESCO-1 specific chimeric antigen receptor gene and a slow virus containing the truncated spleen tyrosine kinase Syk gene. By the combined action of the NY-ESCO-1 specific chimeric antigen receptor gene and the truncated spleen tyrosine kinase Syk gene, the natural killer cell modified by the specific chimeric antigen receptor gene improves the targeted tumor cell specific lethality of the natural killer cell, improves the ability of activating downstream signals of the natural killer cell, avoids the escape of tumor immunity, and accordingly enhances the killing activity on the tumor cell.

Description

Technical field [0001] The present invention relates to the technical field of biopharmaceuticals, in particular to a natural killer cell modified by a specific chimeric antigen receptor gene and its preparation method and application. Background technique [0002] Natural killer cells (NK cells) are the core component of human innate immunity and are the third group of lymphocytes alongside T and B cells. NK cells have a strong non-specific ability to directly kill target cells, and this natural killing activity does not require pre-sensitization by antigens, nor the participation of antibodies, and there is no MHC restriction. NK cells recognize infected and malignant cancerous cells through a variety of cell surface receptors (such as NKG2D, CD16) and natural cytotoxic receptors (NKp44, NKp46, NKp30). These receptors activate the regulatory proteins DAP10 and DAP12 containing the immunoreceptor tyrosine activation motif (ITAM) structure. ITAM can initiate the release of cytot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/867A61K35/17A61P35/00
CPCA61K35/17C12N5/0646C12N9/12C12N15/86C12N2510/00C12N2740/15043C12N2800/22C12Y207/10002
Inventor 不公告发明人
Owner SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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