Chimeric antigen receptor modified dendritic cell containing anti-mesothelin single-chain antibody, and purpose thereof

A chimeric antigen receptor and dendritic cell technology, applied in the field of tumor biological products, can solve the problems of affecting the efficacy of cellular immunity, easy recurrence, and limited efficacy

Active Publication Date: 2017-10-24
SHILI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, CAR-T can produce certain off-target toxicity, which affects its cellular immune efficacy; more importantly, due to the heterogeneity of tumors, there are often multiple tumor antigens in the same tumor, so the efficacy of CAR-T targeting a single tumor antigen Limited and prone to recurrence

Method used

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  • Chimeric antigen receptor modified dendritic cell containing anti-mesothelin single-chain antibody, and purpose thereof
  • Chimeric antigen receptor modified dendritic cell containing anti-mesothelin single-chain antibody, and purpose thereof
  • Chimeric antigen receptor modified dendritic cell containing anti-mesothelin single-chain antibody, and purpose thereof

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Experimental program
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Embodiment

[0058] 1. Determination of mesoCAR gene sequence

[0059] 1.1 The human GM-CSF signal peptide gene SEQ ID NO.3 and the human CD8α hinge region gene SEQ ID NO. .4. Human CD40 transmembrane region and intracellular signal region gene sequence SEQ ID NO.5. The gene sequence of the anti-mesothelin single-chain antibody (anti-SS1scFv) is SEQ ID NO.6.

[0060] 1.2 The above gene sequences were sequentially connected according to the human GM-CSF signal peptide gene, anti-SS1scFv, human CD8α hinge region gene, human CD40 transmembrane region and intracellular signal region sequence to form the final complete GM-CSF-SS1scFv-CD8α- CD40 (mesoCAR) gene sequence SEQ ID NO.2.

[0061] 2. Construction of pCDH-mesoCAR expression plasmid

[0062] 2.1 Whole gene synthesis:

[0063] The complete mesoCAR gene sequence was synthesized by Shanghai Sangon, and restriction sites Xba I / EcoR I were added at both ends.

[0064] 2.2 Cloning the above sequence into pCDH-CMV-MCS-EF1-copGFP-T2A-puro (...

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Abstract

The invention discloses a chimeric antigen receptor, a gene and recombinant expression vector of the chimeric antigen receptor, an engineering mesothelin targeting dendritic cell (mesoCAR-DC) and a purpose thereof. The chimeric antigen receptor (mesoCAR) is GM-CSF-SS1scFv-CD8alpha-CD40, and is formed by connecting human GM-CSF signal peptide, anti-mesothelin single-chain antibodies (anti-SSiscFv), human CD8 alpha hinge regions, human CD40 transmembrane regions and intracellular signal regions. When the dendritic cell modified by the chimeric antigen receptor mesoCAR is used for treating the developing period mesothelin positive cancer (particularly the pancreatic cancer), good specific killing activity is realized on the cancer cells.

Description

technical field [0001] The present invention belongs to the field of tumor biological products, and specifically relates to a chimeric antigen receptor mesoCAR in adoptive immunotherapy and its gene and recombinant expression vector, engineered mesothelin-targeted DC cells (mesoCAR-DC) and its application. Background technique [0002] Dendritic cells (DCs) are a type of mature cells with many dendritic projections that can recognize, ingest and process exogenous antigens and present antigenic peptides to naive T cells and induce T cell activation and proliferation. The most powerful antigen-presenting cells. [0003] The current DC cell immunotherapy is to isolate the tumor patient's own monocytes, culture, expand and induce them to generate DCs in vitro, and then let the DCs load the corresponding tumor antigens, and prepare DCs loaded with tumor antigens after strict screening , and then reinfuse these DC cells back into the corresponding patients to stimulate and activ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/867C12N5/10A61K35/15A61P35/00
Inventor 樊克兴卫莹
Owner SHILI BIOTECH CO LTD
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