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PD1-knockout chimeric antigen receptor T cell targeting CD20, preparation method and application thereof

A chimeric antigen receptor, CAR-CD20 technology, applied in the field of medical biology, can solve the problem that CART-CD19 can no longer exert anti-tumor effect

Inactive Publication Date: 2019-06-04
SHENZHEN BINDEBIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since tumors of the same type often express different immune phenotypes, for example, when tumor cells escape CD19-negative, that is, there are tumor cells that do not express CD19, the CD19 target will fail, and CART-CD19 will no longer be able to function. Anti-tumor effect

Method used

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  • PD1-knockout chimeric antigen receptor T cell targeting CD20, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting CD20, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting CD20, preparation method and application thereof

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Embodiment 1

[0091] A method for preparing CD20-targeted chimeric antigen receptor T cells knocked out of PD1, comprising the following steps:

[0092] (1) Prepare the gene sequence of chimeric antigen receptor CAR-CD20

[0093] Prepare the coding genes of signal peptide, single-chain antibody targeting CD20, CD8α hinge region, CD8 transmembrane region, 4-1BB signal region and CD3ζ signal region respectively, the coding gene of the signal peptide is shown in SEQ ID NO: 8 , the coding gene of the single-chain antibody targeting CD20 is shown in SEQ ID NO: 5, the coding gene of the CD8α hinge region is shown in SEQ ID NO: 10, and the coding gene of the CD8 transmembrane region is shown in SEQ ID NO: As shown in ID NO: 12, the coding gene of the 4-1BB signal region is shown in SEQ ID NO: 14, and the coding gene of the CD3ζ signal region is shown in SEQ ID NO: 16.

[0094] The genes encoding the above signal peptide, CD20-targeting single-chain antibody, CD8α hinge region, CD8 transmembrane r...

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Abstract

The invention provides a preparation method of a PD1-knockout chimeric antigen receptor T cell targeting CD20. The method comprises: linking an encoding gene of the chimeric antigen receptor CAR-CD20and targeting CD20 to a vector, and constructing the recombinant lentivirus, performing transfection on CD3 positive T lymphocytes, and performing knockout of the PD1 gene of the transfected T cells to obtain the PD1-knockout chimeric antigen receptor T cell targeting CD20. The knockout of the PD1 gene by the T cell is in favor of amplification of T cells in the patient, and avoids the escape of immunosurveillance of the tumor cells, and the T cell has the performance of killing CD20 positive tumor cells with high efficiency and specificity, which maintains the viability and lethality of the cells for a long time.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a chimeric antigen receptor T cell targeting CD20 knocked out of PD1 and its preparation method and application. Background technique [0002] CAR-T (Chimeric Antigen Receptor T Cell) technology is a new type of cell therapy, which is to reinfuse CAR-modified T cells into the human body, activate the autoimmune system, and kill tumor cells. The treatment of non-Hodgkin's lymphoma has a remarkable curative effect and is considered to be one of the most effective treatments for malignant tumors. [0003] At present, CAR-T technology has achieved remarkable results in CD19-positive blood tumors (such as B-cell lymphoma, acute and chronic B-lymphoblastic leukemia). However, since tumors of the same type often express different immune phenotypes, for example, when tumor cells escape CD19-negative, that is, there are tumor cells that do not express CD19, the CD19 target will fail, and C...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N7/01C12N15/113C12N5/10A61P35/00
Inventor 曾滢张长风吴咏东
Owner SHENZHEN BINDEBIOTECH CO LTD
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