Immune cell modified by suppressor protein blocking type chimeric antigen receptor and application of immune cell

A chimeric antigen receptor, immune cell technology, applied in genetically modified cells, cells modified by introducing foreign genetic material, blood/immune system cells, etc., can solve problems such as enhancing the anti-tumor activity of CAR-T cells, Achieve the effect of enhancing anti-tumor immune response, accurately determining the dosage, and reducing the cost of medication

Pending Publication Date: 2022-01-11
苏州璞惠卓越生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no literature disclosing the blocking protein (Blocker) to enhance the anti-tumor activity of CAR-T cells

Method used

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  • Immune cell modified by suppressor protein blocking type chimeric antigen receptor and application of immune cell
  • Immune cell modified by suppressor protein blocking type chimeric antigen receptor and application of immune cell
  • Immune cell modified by suppressor protein blocking type chimeric antigen receptor and application of immune cell

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Experimental program
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Embodiment 1

[0031] (1) Construct the expression plasmid of CAR-Blocker

[0032] CAR is a CAR molecule targeting CD19, the sequence is SEQ ID NO: 1-2 in Table 1, the sequence of blocking protein Blocker is SEQ ID NO: 3-4, CAR and blocking protein Blocker are connected by self-cleaving polypeptide linker sequence 2A, so that CAR and Blocker can be co-expressed to obtain an arrestin-blocking chimeric antigen receptor CAR-Blocker, and the sequence of CAR-Blocker is SEQ ID NO: 5-6. The viral expression plasmid (AddgeneID: #12257) used was digested with BamHI / NdeI as the backbone, and the CAR-Blocker was synthesized and inserted into the vector.

[0033] (2) Preparation of lentivirus

[0034] A three-plasmid packaging method was adopted, including an expression plasmid, a packaging helper plasmid psPAX2 (AddgeneID: #12260) and pMD2.G (AddgeneID: #12259). The preparation process is as follows: cryopreserved HEK293T cells (purchased from Shanghai Institute of Cell Research, Chinese Academy of S...

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Abstract

The invention relates to an immune cell modified by a suppressor protein blocking type chimeric antigen receptor and application of the immune cell. The immune cell modified by the suppressor protein blocking type chimeric antigen receptor is used for a medicine for treating malignant tumors, and the suppressor protein and the chimeric antigen receptor are co-expressed in the immune cell; the nucleotide sequence of the suppressor protein is as shown in SEQ ID NO: 3, and the protein sequence is as shown in SEQ ID NO: 4; and the nucleotide sequence for blocking co-expression of the suppressor protein and the chimeric antigen receptor is as shown in SEQ ID NO: 5, and the protein sequence is as shown in SEQ ID NO: 6. From the perspective of blocking transduction of a negative regulation signal of the immune cell, the CAR-T cell expresses the suppressor protein, the transmission of the negative regulation signal is weakened / blocked, and the anti-tumor activity of the CAR-T cell is enhanced; and the anti-tumor immune response can be enhanced without being combined with an immune checkpoint antibody drug, the drug cost is reduced, the dosage can be accurately determined, and the curative effect is exact and remarkable.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an immune cell modified with an inhibitory protein-blocking chimeric antigen receptor and its application. Background technique [0002] At present, CAR-T immunotherapy has made remarkable clinical progress in the field of tumor treatment, and immune checkpoint blockers such as PD1 / PD-L1 antibody drugs have also achieved gratifying curative effects in tumor treatment. CAR-T immunotherapy is to extract immune cells, expand them through in vitro culture, and then use genetic engineering technology to inlay immune cells with gene fragments that can recognize cancer cells, and then reinfuse the immune cells into the patient's body; PD- 1 Antibody drug itself will not kill cancer cells, it reactivates immune cells by releasing the inhibition of immune cells; the anti-tumor mechanism of the two is different, in order to further improve the curative effect, clinical research has carried out C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/62A61K39/00A61P35/00
CPCC12N5/0636C12N5/0646C07K16/2803C12N9/16C12Y301/03048A61K39/001112A61P35/00C12N2510/00C07K2319/33C07K2319/00A61K2039/5156
Inventor 杨光朱毅敏
Owner 苏州璞惠卓越生物科技有限公司
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