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A chimeric antigen receptor NK cell and a preparation method and an application thereof

A chimeric antigen receptor, NK cell technology, applied in receptors/cell surface antigens/cell surface determinants, biochemical equipment and methods, animal cells, etc., can solve the problem of killing tumor cells to be further improved, T cell separation Difficulty in purification, graft-versus-host reaction and other problems, to achieve the effect of enhancing killing effect, improving anti-tumor activity, and powerful function

Inactive Publication Date: 2019-01-15
EAST CHINA NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, problems such as off-target effects, cytokine storm, difficulty in separation and purification of T cells, low infection efficiency, graft-versus-host reaction, and long production cycle in CAR-T therapy are still waiting for us to solve
[0004] However, at present, the effect of killing tumor cells with only CAR single-modified effector cells still needs to be further improved

Method used

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  • A chimeric antigen receptor NK cell and a preparation method and an application thereof
  • A chimeric antigen receptor NK cell and a preparation method and an application thereof
  • A chimeric antigen receptor NK cell and a preparation method and an application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0040] Preparation of chimeric antigen receptor NK cells co-expressing IL-15 targeting CD19 extracellular region

[0041] 1 Construction of lentiviral packaging plasmid

[0042] The amino acid sequence of scFv targeting the extracellular domain of CD19 is shown in SEQ ID NO.1.

[0043] Link the nucleotide sequence of the chimeric antigen receptor targeting the extracellular segment of CD19 with the nucleotide sequence of IL-15 through IRES to obtain the co-expressed nucleic acid sequence SEQ ID NO.7, and clone SEQ ID NO.7 into pCDH vector (for structure see Figure 5 ), the restriction sites are on SalI and XbaI. The structural positional relationship of each expression element is as follows figure 1 shown.

[0044] The nucleotide sequence of the chimeric antigen receptor targeting the extracellular segment of CD19 sequentially includes: single-chain antibody scFv, CD8α transmembrane domain, 4-1BB co-stimulatory signal transduction region and CD3ζ signal transduction domai...

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PUM

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Abstract

The invention discloses a chimeric antigen receptor NK cell, a preparation method and an application thereof, which relate to the technical field of cellular immunotherapy. The chimeric antigen receptor NK cell disclosed by the invention can simultaneously express IL-15 and a chimeric antigen receptor targeting CD19. The chimeric antigen receptor NK cells have good killing effect on tumor cells, and can be used for preparing anti-tumor drugs, thus providing a new idea and method for treating tumor.

Description

technical field [0001] The invention relates to the technical field of cellular immunotherapy, in particular to a chimeric antigen receptor NK cell and its preparation method and application. Background technique [0002] In July 2017, the FDA Oncology Drug Expert Advisory Committee unanimously approved and approved the launch of Novartis' CAR-T therapy CTL-019. To the "cusp". CTL019, as the first chimeric antigen receptor T cell (CAR-T) product approved for marketing, marks the arrival of a new era of tumor immunotherapy. [0003] While CAR-T brings hope to the vast number of tumor patients, it also brings many treatment risks. At present, problems such as off-target effects, cytokine storm, difficulty in separation and purification of T cells, low infection efficiency, graft-versus-host reaction, and long production cycle in CAR-T therapy are still waiting for us to solve. [0004] But at present, the tumor cell killing effect of effector cells only modified by CAR stil...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/62A61K35/17A61P35/00
CPCA61P35/00A61K35/17C12N5/0646C07K14/7051C07K2319/03C12N2510/00
Inventor 江文正牛聪义韦欢李贞龙刘明耀席在喜
Owner EAST CHINA NORMAL UNIVERSITY
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