Natural killer cell modified by specific chimeric antigen receptor gene and preparation method and use thereof

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: 2018-04-10
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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  • Natural killer cell modified by specific chimeric antigen receptor gene and preparation method and use thereof
  • Natural killer cell modified by specific chimeric antigen receptor gene and preparation method and use thereof
  • Natural killer cell modified by specific chimeric antigen receptor gene and preparation method and use thereof

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preparation example Construction

[0057] The present invention also proposes a method for preparing natural killer cells modified by specific chimeric antigen receptor genes, which is used for preparing the natural killer cells modified by specific chimeric antigen receptor genes. Wherein, the preparation method comprises the following steps:

[0058] Infect NK cells simultaneously 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 one embodiment, the first lentivirus is obtained as follows:

[0060] S11: mixing 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 embedded in 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 carrying the NY-ESO-1 / scFv-CD8-41BB-DA...

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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 invention relates to the technical field of biopharmaceuticals, in particular to a specific chimeric antigen receptor gene-modified natural killer cell 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 does it require the participation of antibodies, and there is no MHC restriction. NK cells recognize infected and malignant cells through various cell surface receptors (eg NKG2D, CD16) and natural cytotoxicity receptors (NKp44, NKp46, NKp30). These receptors activate regulatory proteins DAP10, DAP12 that contain immunoreceptor tyrosine activation motif (ITAM) structures, ITAMs can initiate the release of cytotoxic granule pr...

Claims

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Application Information

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Patent Type & Authority Applications(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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