A kind of target cell for testing car-t cell killing effect and its preparation method and application

Abstract

The present invention provides a target cell CCC-3T3 for testing the killing effect of CAR-T cells, which belongs to the technical field of cell engineering; the target cell CCC-3T3 expresses CAR-T cell target antigen genes and Caveolin-1 and caspase ‑3 fusion protein gene in mouse fibroblast-like cells. The preparation method of the target cell CCC-3T3 is: using the lentiviral transfection method to transfer the fusion protein gene of Caveolin-1 and caspase-3 into mouse fibroblast-like cells to obtain CC-3T3 cells expressing the fusion protein; Then, the CAR-T cell target antigen gene was transferred into CC-3T3 cells by lentiviral transfection method to obtain target cells CCC-3T3. The target cell CCC-3T3 has a low killing background, is sensitive to killing signals, and can accurately reflect the specific killing ability of effector cells.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and in particular relates to a target cell for testing the killing effect of CAR-T cells, a preparation method and application thereof. Background technique [0002] The killing function is an important aspect of the body's immune function. There are a variety of target cells with killing functions in the immune system, such as natural killer cells (NK), cytotoxic T cells (CTL), monocytes and macrophages with ADCC function Wait. The full name of CAR-T cells is Chimeric Antigen Receptor T cells. It is one of the more effective treatments for malignant tumors. Similar to other immunotherapies, its basic principle is to use the patient's own immune cells to eliminate cancer cells. [0003] The target cells of conventional killing experiments mostly use human immortalized cells or tumor cells, such as K562 cells, 293T cells, etc. Since these cells are human immortalized cells, many mutant p...

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Problems solved by technology

[0003] The target cells of conventional killing experiments mostly use human immortalized cells or tumor cells, such as K562 cells, 293T cells, etc. Since these cells are human immortalized cells, many mutant proteins other than the target antigen protein will be expressed on the cell surface , or presented to the cell surface by HLA molecules, which causes effector cells to kill target cells, in addition to the specific killing of target antigens, there is also non-specific killing of other proteins, and the killing background is high. When the effector cells When the proportion of CAR-T cells in specific T cells is low, false negative results will appear

In addition, conventional target cells are not sensitive to killing signals, and their self-proliferation speed is too fast, so they cannot be used for long-term killing experiments.

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