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Application of NAD < + > metabolic targeting-based NK cells in disease treatment

A technology of NK cells and target cells, which is applied in the fields of biomedicine and molecular biology, can solve the problems that solid tumors cannot achieve satisfactory results, achieve good application prospects, important development value, and improve the ability to kill target cells

Pending Publication Date: 2022-05-13
SHANDONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, although the application of NK cells expanded in vitro to treat a variety of non-solid tumors has achieved certain curative effects, it has not yet achieved satisfactory results for solid tumors.

Method used

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  • Application of NAD &lt; + &gt; metabolic targeting-based NK cells in disease treatment
  • Application of NAD &lt; + &gt; metabolic targeting-based NK cells in disease treatment
  • Application of NAD &lt; + &gt; metabolic targeting-based NK cells in disease treatment

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Experimental program
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Effect test

Embodiment

[0079] 1. Acquisition and cultivation of human NK cells:

[0080] 1. NK-92 cell line: purchased from ATCC cell bank, cultured in α-MEM+12.5%FBS+12.5%HBS+0.2mM inositol+0.1mM 2-mercaptoethanol+0.02mM folic acid+100U / ml recombinant human IL In the complete medium of -2, subculture once every 3 days.

[0081] 2. Purification and cultivation of human peripheral blood NK cells: extract 10ml of healthy human peripheral blood, use the magnetic bead purification kit to sort NK cells, and culture them in α-MEM+12.5%FBS+12.5%HBS+0.2mM inositol+0.1 mM 2-mercaptoethanol + 0.02mM folic acid + 100U / ml recombinant human IL-2 complete medium for subsequent experiments.

[0082] 2. The activation of NK cells is accompanied by an increase in the metabolic level of NAD+:

[0083] Stimulation and activation of NK cells by cytokine IL-12 combined with IL-15 or Poly(I:C) can significantly increase their intracellular NAD+ and NAD / NADH levels ( figure 1 A), and intracellular NAD + The expression...

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Abstract

The invention provides application of NK cells based on NAD < + > metabolism targeting in disease treatment, and belongs to the technical field of biological medicine and molecular biology. Researches find that along with the increase of the NAD + metabolism level in the NK cell activation process, the NAD + level in the NK cells can be improved by supplementing the NAD + precursor molecules, and the NAD + can remarkably improve the survival ability of the NK cells and the cytokine secretion and target cell killing ability of the NK cells, so that the anti-tumor and anti-infection effects of the NK cells are enhanced. A new thought is provided for application of NK cell transfection in treatment of diseases such as tumors and chronic infection, and important development value and good application prospects are achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and molecular biology, in particular to NAD-based + Metabolically targeted NK cells have applications in disease therapy. Background technique [0002] The information disclosed in the Background of the Invention is only intended to increase the understanding of the general background of the invention, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] So far, diseases such as malignant tumors and chronic infections are still important causes of harm to human health. In recent years, improving the immune response ability of patients through active or passive means has become an important means of clinical combined treatment of related diseases. In particular, immune checkpoint molecule (PD-1, CTLA-4, etc.) inhibitors, CAR-T cells and other therapeutic meth...

Claims

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Application Information

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IPC IPC(8): A61K31/706A61K31/455A61K31/47A61K35/17A61P35/00A61P31/00
CPCA61K31/706A61K31/455A61K31/47A61K35/17A61P35/00A61P31/00A61K2300/00
Inventor 梁晓红郭小维马春红高立芬谭思雨李春阳武专昌
Owner SHANDONG UNIV
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