Recombinant lentiviral expression vector, reconstitution cell and application of reconstitution cell to culturing natural killer cell

Abstract

The invention provides a recombinant lentiviral expression vector, and belongs to the technical field of cell engineering. The recombinant lentiviral expression vector comprises a 21mb gene, a 41BBL gene and an MICA gene which are sequentially connected in series; and the 21mb gene is obtained by sequentially connecting a signal peptide gene, an IL21 gene, a CD8 Hinge gene and a CD8 transmembranegene in series. After the provided recombinant lentiviral expression vector transfects a host cell, the host cell expresses the MICA and the 41BBL, and the IL21 is fused and expressed (but not secreted in a culture environment) on a cytomembrane of the host cell. After the provided recombinant lentiviral expression vector transfects the host cell, an obtained reconstitution cell promotes the highexpression of the NKG2D, avoids excessive activation of an NK cell and prolongs the proliferation time of the NK cell in the process of induction culture of the NK cell.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and in particular relates to a recombinant lentiviral expression vector, recombinant cells and their application in natural killer cell culture. Background technique [0002] Natural killer cells (NK) are important immune cells in the body, not only related to anti-tumor, anti-viral infection and immune regulation, but also involved in the occurrence of hypersensitivity and autoimmune diseases in some cases, and can recognize Target cells, killing mediators. The exact source of natural killer cells is not very clear, generally believed to be derived directly from the bone marrow. In view of the wide application of natural killer cells, providing an efficient and stable method for inducing natural killer cells has broad application prospects. [0003] The Chinese invention patent with the application number CN201710096536.1 discloses a method of efficiently expanding NK cells using K562...

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Problems solved by technology

[0003] The Chinese invention patent with the application number CN201710096536.1 discloses a method of efficiently expanding NK cells using K562 engineered cells with high expression of membrane proteins CD19, CD137L, CD86, CD64 and transmembrane protein IL-21 combined with human IL-2 mutants method, although this method solves the problem of NK cells specifically recognizing CD19+ cells, it cannot effectively stimulate and improve the expression of NKG2D receptors that are crucial for NK cell tumor killing

The Chinese invention patent with the application number CN201310173235.6 discloses a method for expanding and activating lymphocytes and NK cells using CD8α-interleukin 21 fragment-CD137 transfected K562 cells. This method has a low level of cell activation and limits cell proliferation Trend and lethality

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