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Universal cell therapy product for expressing antigen recognition regions, and preparation method and application thereof

An antigen recognition and cell technology, applied in the field of biomedicine, can solve problems such as adverse reactions, restrictions on the wide application of CART, and general-purpose cell therapy products that do not express antigen recognition regions, so as to reduce the burden, reduce cell preparation and transportation costs , the effect of shortening the cell preparation cycle

Active Publication Date: 2019-10-15
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

Although T cells play a key role in CART therapy, it is also the reason that limits the wide application of CART and causes many adverse reactions
[0004] There is no general-purpose cell therapy product targeting the expression antigen recognition region of leukemia

Method used

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  • Universal cell therapy product for expressing antigen recognition regions, and preparation method and application thereof
  • Universal cell therapy product for expressing antigen recognition regions, and preparation method and application thereof
  • Universal cell therapy product for expressing antigen recognition regions, and preparation method and application thereof

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Embodiment 1

[0033] 1 Cell preparation

[0034] According to the CTL019 sequence, the present invention designs CD19CAR and CD19 antigen recognition regions, and obtains the heavy chain (VH) and light chain (VH) and light chain ( VL) cDNA sequence, spliced ​​and synthesized to bind to the CD133 antigen recognition region, introduce membrane localization signal (CD8leader), hinge region (CD8 hinge), transmembrane region (CD8 transmenber), co-stimulatory signal (CD28, 4-1BB, CD3- zata) (see Table 1 for the sequence). Entrust Sangon Bioengineering (Shanghai) Co., Ltd. to synthesize the entire expression frame, amplify it by PCR, introduce restriction sites EcoRI and BamHI, and insert it into the pCDH-MCS-T2A-copGFP-MSCV virus vector ( Figure 7 , purchased from System Biosciences, Cat. No. CD523A-1) Between the EcoRI-BamHI sites, after the sequencing was correct, the plasmid was extracted using the plasmid extraction kit from Promega. Lentiviral packaging: using pSPAX2, pMD2G viral packagin...

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Abstract

The present invention relates to a universal cell therapy product for expressing antigen recognition regions, and a preparation method and an application thereof. The antigen recognition regions aiming at tumor cell antigens are carried on universal cells without an HLA restriction, the antigens are degraded after treatment, and tumor cells are changed in series until death, thereby playing a rolein treatment. A provided new strategy for treating tumors is expected to replace individualized CART, remarkably reduces preparation costs of CART cells, achieves an aim of treating the tumors, doesnot generate cytokine storm and neurotoxicity, reduces threshold of CART treatment, and benefits more patients.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a general-purpose cell therapy product expressing an antigen recognition region and its preparation method and application. Background technique [0002] Chimeric Antigen Receptor T cells (CART) are genetically modified so that the single-chain variable region (scFv) of a monoclonal antibody that can specifically recognize the target antigen is expressed on the surface of T cells, and at the same time The scFv is coupled to the artificially designed T cell intracellular activation and proliferation signal threshold through the transmembrane region. Therefore, the specific recognition of the target antigen by the monoclonal antibody is combined with the function of the T cell to produce a specific killing effect, and CART can kill the target cell in a non-MHC-restricted manner. B-cell non-Hodgkin's lymphoma (B-NHL) tumor cells stably express CD19 molecule, which provides a ta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867A61K39/00A61P35/02
CPCC12N5/0662C07K16/2803C12N15/86A61K39/001102A61P35/02C12N2510/00C07K2317/622C12N2740/15043A61K2039/5156Y02A50/30
Inventor 刘晗李丹王睿恒许琼予谢树峰
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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