Target cell TCC-3T3 for inspecting killing effect of TCR-T (T cell receptor T) cell as well as preparation method and application thereof

Abstract

The invention provides a target cell TCC-3T3 for inspecting killing effect of TCR-T (T cell receptor T) cell, and belongs to the technical field of cell engineering. The target cell TCC-3T3 is a mousefibroblast-like cell for expressing a target antigen gene of the TCR-T cell, an HLA (human leukocyte antigen) gene, and a fusion protein gene of Caveolin-1 and caspase-3. A preparation method of thetarget cell TCC-3T3 comprises the 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 toobtain the cell CC-3T3 for expressing the fusion protein; transferring the target antigen gene of the TCR-T cell and the HLA gene into the cell CC-3T3 by the lentiviral transfection method, so as to obtain the target cell TCC-3T3. The target cell TCC-3T3 has the advantages that the killing background is low; the target cell TCC-3T3 is sensitive to the killing signals; the specific killing abilityof effector 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 TCR-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. TCR-T cells are T cells genetically engineered by the receptor, and the adoptive immunotherapy method for tumors based on modified T cells can exert a strong anti-tumor effect in vivo by virtue of its high affinity recognition ability for tumor-specific antigens. Tumor immune effect: 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 c...

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 specific T cells TCR-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.

Images