Technology for preparing universal humanized CAR19-DNT cell and application of universal humanized CAR19-DNT cell

Abstract

The invention provides a technology for preparing a universal humanized CAR19-DNT cell and application of the universal humanized CAR19-DNT cell. Specifically, the invention provides a universal CAR-T cell targeting CD19, the universal CAR-T cell expresses an exogenous CAR construct, the CAR construct has a structure as shown in a formula I, L-scFv-H-TM-C-CD3 zeta, in the formula I, L is none or a signal peptide sequence, the scFv is an antibody single-chain variable region sequence targeting CD19; H is a null or hinge region; TM is a transmembrane domain; C is a costimulatory signal molecule; CD3 zeta is a cytoplasm signaling sequence derived from CD3 zeta. The universal humanized CAR19-DNT cell has the advantages of low immunogenicity, no need of gene editing to avoid GvHD, high safety, high specificity and remarkable tumor killing effect, and the construction method provided by the invention can realize large-scale production and is low in production cost.

Description

technical field [0001] The invention belongs to the technical field of tumor immune cell therapy, and specifically relates to a technical method for preparing general-purpose humanized antigen chimeric receptor CART-19 cells and an application thereof. Background technique [0002] Chimeric Antigen Receptor T (CART) cells are currently one of the most promising tumor immunotherapies. The two CART cell products approved by the US FDA in 2017 are isolated from the peripheral blood of tumor patients themselves. Mononuclear cells (Peripheral Blood Mononuclear Cell, PBMC) cells are embedded with specific antigen receptors (CARs) through genetic engineering, which enhances the targeting, killing activity and persistence of T cells in PBMCs, and its ability to detect tumor cell surface antigens Recognition of MHC-independent constraints. A common CAR consists of an extracellular antigen-binding region, a transmembrane region, and an intracellular T cell receptor signal transductio...

AI Technical Summary

Problems solved by technology

[0004] Although the effect of targeting CD19 CART in clinical trials is very significant, there are still many defects. In addition to causing severe side effects such as cytokine storm, there are 4 major problems: First, some lymphocytes have low quantity or poor quality Advanced patients lost the chance of CART treatment due to lack of sufficient number of autologous immune cells; secondly, it has been reported that when CART products prepared from patients' autologous cells were used, viral genes were transduced into residual tumor cells in peripheral blood, resulting in patient death; moreover, most of the general-purpose CART cell products currently under development use gene editing technology to knock out genes that can cause graft-versus-host disease, such as TCR receptors, etc., but knocking out these genes requires a cumbersome construction process And large-scale sequencing, and the off-target rate is high; finally, because the preparation of autologous CART cell products is a 1-to-1 individualized treatment, the preparation cost is expensive, and the preparation process requirements are complicated, resulting in high product prices, which are unaffordable for most patients and increase social medical burden.

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