Target cell for inspecting killing effect of CAR-T (chimeric antigen receptor T) cell as well as preparation method and application thereof

Abstract

The invention provides a target cell CCC-3T3 for inspecting killing effect of CAR-T (chimeric antigen receptor T) cell, and belongs to the technical field of cell engineering. The target cell CCC-3T3is a mouse fibroblast-like cell for expressing a target antigen gene of the CAR-T cell and a fusion protein gene of Caveolin-1 and caspase-3. A preparation method of the target cell CCC-3T3 comprisesthe following steps of transferring the fusion protein gene of Caveolin-1 and caspase-3 into the mouse fibroblast-like cell by a lentiviral transfection method, so as to obtain the cell CC-3T3 for expressing the fusion protein; transferring the target antigen gene of the CAR-T cell into the cell CC-3T3 by the lentiviral transfection method, so as to obtain the target cell CCC-3T3. The target cellCCC-3T3 has the advantages that the killing background is low; the target cell CCC-3T3 is sensitive to the killing signals; the specific killing ability of effect cells can be accurately reflected.

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, and a preparation method and application thereof. Background technique [0002] Killing function is an important aspect of the body's immune function. There are a variety of target cells with killing function in the immune system, such as natural killer cells (NK), cytotoxic T cells (CTL), monocytes and macrophages with ADCC function. Wait. CAR-T cells, called chimeric antigen receptor T cells, are currently one of the more effective treatments for malignant tumors. Similar to other immunotherapies, the basic principle is to use the patient's own immune cells to eliminate cancer cells. [0003] Conventional killing experiment target cells, mostly human immortalized cells or tumor cells, such as K562 cells, 293T cells, etc., because these cells are human immortalized cells, many mutant proteins other than ...

AI Technical Summary

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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