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PTPN2-blocked immune cell and application and preparation thereof

A technology of immune cells and NKT cells, applied in the field of PTPN2-blocking immune cells and their applications and preparations, can solve the problems of solid tumors without good treatment effect, low clinical response rate, off-target, etc., to enhance the killing function and reduce exhaustion. , to avoid toxic effects

Inactive Publication Date: 2020-12-22
GUANGDONG XIANKANGDA BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although CAR-T cell therapy shines in the treatment of hematological tumors, it does not have a good therapeutic effect on solid tumors. T cells are not easy to enter the tumor, cannot expand in large quantities in the tumor, and are prone to off-target and other toxic side effects. is a major contributor to its low clinical response rate

Method used

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  • PTPN2-blocked immune cell and application and preparation thereof
  • PTPN2-blocked immune cell and application and preparation thereof
  • PTPN2-blocked immune cell and application and preparation thereof

Examples

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Effect test

Embodiment 1

[0080] The preparation method and functional verification of immune cells in this embodiment include the following steps:

[0081] Step 1: Isolation of PBMC from peripheral blood and expansion of T cells

[0082] Mononuclear cells were isolated from donor peripheral blood, density gradient centrifugation was performed using ficol, and T cells were enriched with a T cell sorting kit (CD3 MicroBeads, human-lyophilized, 130-097-043), using conjugated anti-CD3 / anti-CD28 magnetic beads activated culture and expansion of T cells; the medium used TexMACS GMP Medium (MiltenyiBiotec, 170-076-309), containing 10% FBS, 2mM L-glutamine, 100IU / ml rhIL2, all cells were placed Cultured in a constant temperature incubator at 37°C with 5% CO2.

[0083] Step 2: Cell Line Culture

[0084] Cell line expressing GPC3: Huh-7 (human liver cancer cell), purchased from ATCC.

[0085] Cell line not expressing GPC3: A549 (human non-small cell lung cancer cells), purchased from ATCC.

[0086] Cells f...

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Abstract

The invention relates to the field of biological cells, and particularly relates to a PTPN2-blocked immune cell and application and a preparation thereof. The PTPN2-blocked immune cell expresses a chimeric antigen receptor, the PTPN2 function in the immune cell is blocked, and the immune cell can secrete an anti-PD-1 antibody. The invention provides a method for killing tumors and reversing exhaustion. The PTPN2-blocked function is innovatively combined with CAR and anti-PD-1 immunoregulation, so that the tumor killing activity of the immune cell is improved, and the effect of curing tumors isachieved.

Description

technical field [0001] The present invention relates to the technical field of biological cells, in particular to an immune cell that blocks PTPN2 and its application and preparation, an immune cell that expresses CAR, blocks the function of intracellular PTPN2 protein, and optimizes the expansion ability and killing function of CD8+ T cells. And combined with the PD-1 inhibitory pathway, the immunotherapy response was more pronounced. Background technique [0002] In recent years, tumor immunotherapy has developed rapidly, especially adoptive cell therapy (ACT), which means that immune cells such as T cells and NK cells are isolated from patients, modified and expanded in vitro, and then reintroduced into patients for tumor treatment. method of treatment. In 2013, the immunotherapy of tumors was rated as the first among the "Top Ten Breakthroughs" by Science Impurities. [0003] CAR-T and TCR-T are important components of adoptive cell therapy, especially CAR-T therapy, w...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867C12N15/62C12N15/13A61K39/00A61P35/00
CPCA61K2039/5156A61P35/00A61K39/001102C07K14/7051C07K16/2818C07K16/303C07K16/40C07K2317/622C07K2319/02C07K2319/03C07K2319/33C07K2319/74C12N5/0636C12N15/86C12N2510/00C12N2740/15043C12N2800/107
Inventor 马丽雅贾晓晓方晓
Owner GUANGDONG XIANKANGDA BIOTECH CO LTD
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