COMPOSITION COMPRISING NK CELLS AND plant-derived EXOSOMES, USE THEREOF AND PRODUCTS COMPRISING SAME

A technology derived from NK cells and plants, applied in the field of extracellular vesicle compositions, can solve the problems of enhancing the ability of NK cells to attack cancer cells and reducing the ability to attack cancer cells, so as to overcome the reduced ability of cancer cells and enhance the ability to attack , enhance the effect of treatment

Pending Publication Date: 2021-10-01
上海瑞开投资管理有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Plant-derived extracellular vesicles can activate the release of NK cell perforin and granzyme, enhance the ability of NK cells to attack cancer cells, and overcome the previous problem that NK cells themselves have reduced ability to attack cancer cells

Method used

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  • COMPOSITION COMPRISING NK CELLS AND plant-derived EXOSOMES, USE THEREOF AND PRODUCTS COMPRISING SAME
  • COMPOSITION COMPRISING NK CELLS AND plant-derived EXOSOMES, USE THEREOF AND PRODUCTS COMPRISING SAME
  • COMPOSITION COMPRISING NK CELLS AND plant-derived EXOSOMES, USE THEREOF AND PRODUCTS COMPRISING SAME

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Experimental program
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preparation example Construction

[0047] In the preparation method of extracellular vesicles described above, the step of "extracting fucoidan from seaweed" can be omitted. In this case, fucoidan was used as a starting material from which extracellular vesicles were extracted.

[0048]In a preferred embodiment, the molecular weight of the fucoidan is less than or equal to about 500 Daltons, such as less than or equal to 400 Dal, less than or equal to 450 Dal, less than or equal to 460 Dal, less than or equal to 470 Dal, less than or equal to 480 Dal, less than or equal to Or equal to 490Dal, less than or equal to 495Dal, less than or equal to 500Dal, less than or equal to 505Dal, or less than or equal to 510Dal.

[0049] In one embodiment of the present application, the extracellular vesicle is an exosome

[0050] NK cells

[0051] The plant-derived extracellular vesicles described in this application can be used as NK cell activators to activate NK cells. In a specific embodiment of the present application...

Embodiment 1

[0095] Example 1: Preparation of plant-derived extracellular vesicles

[0096] 1. Transfer 1 package of power fucoidan (JAN:4580123711060) to a 50mL test tube.

[0097] 2. Use a 20ml syringe (without needle) to go in and out 5 times to make it fully even.

[0098] 3. Separately add 1 g aliquots to 50 mL PBS.

[0099] 4. Stir with a stirrer for 1 hour.

[0100] 5. Let stand for 15 minutes.

[0101] 6. Take 45mL supernatant and centrifuge at 2,000×g, 4°C for 10 minutes.

[0102] 7. Take the supernatant (do not take the sediment).

[0103] 8. Through 0.22um filter.

[0104] 9. Centrifuge at 35,000 rpm for 70 minutes at 4°C.

[0105] 10. Wash the pellet with PBS.

[0106] 11. Centrifuge again at 35,000 rpm at 4°C for 70 minutes.

[0107] 12. Discard the supernatant, and transfer the PBS liquid containing the suspension at the bottom to a 1.5mL test tube.

[0108] 13. Centrifuge at 10,000 x g at 4°C for 10 minutes (if there is a green precipitate, remove it).

[0109] 14....

Embodiment 2

[0110] Example 2: The effect of plant-derived extracellular vesicles on enhancing the content of granzymes in NK cells

[0111] First, human NK cells (Lonza, Poietics TMHuman NK cells; product code: 2W-501) were quantified by the amount of granzyme B, which was set to 1.0. Then, human NK cells were incubated with the plant-derived extracellular vesicles prepared in Example 1 at a concentration of 100 extracellular vesicles / human NK cells for 48 hours, and then the relative value of the amount of granzyme B was calculated. A control group was also set up. The amount of granzyme B was quantified by ELISA method using Granzyme B ELISA Development Kit (Human Alkaline Phosphatase) (product code: 3485-1A-6, Cosmo Bio Inc.) following the protocol of the attached instructions. The experimental results are plotted in figure 1 middle.

[0112] Such as figure 1 As shown, the expression of granzyme B in NK cells increased more than 8-fold after the addition of plant-derived extracell...

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Abstract

The present application provides a composition comprising NK cells and plant-derived extracellular vesicles. The extracellular vesicles can act as NK cell activators to stimulate NK cells to release proteins, such as perforin and / or granzyme, thereby exerting cytotoxicity. The plant-derived extracellular vesicles overcome the problem that the ability of NK cells to attack cancer cells is reduced in the past, enhances the ability of the NK cells to attack the cancer cells, and improves the treatment effect of the NK cells on cancers.

Description

technical field [0001] The application belongs to the field of biomedicine, and specifically relates to a composition comprising NK cells and plant-derived extracellular vesicles, its use and products comprising the same. Background technique [0002] On the surface of natural killer cells (natural killer cells, NK cells), receptor molecules (KIR) that inhibit NK cell activation are expressed. Since major histocompatibility complex (MHC) molecules can bind to KIRs, cells expressing MHC molecules can send signals so that they are not killed by NK cells. Normal cells always express MHC molecules to express themselves, so NK cells will not kill normal cells expressing MHC molecules. If a "cancer cell" that does not express MHC proteins appears, NK cells will be a mechanism to kill it. [0003] NK cell therapy aims to apply this property to cancer treatment. However, experience has shown that treating solid tumors with NK cells is very difficult. One of the reasons is that n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A61K35/17A61P35/00A61P35/02
CPCC12N5/0646A61K35/17A61P35/00A61P35/02C12N2500/76
Inventor 刘冠宏
Owner 上海瑞开投资管理有限公司
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