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Application of CD137 antibody in preparation of drugs for promoting NK cells to express CD16 molecules

A CD137, 1. CD137 technology, applied in the field of medicine, can solve the problems of treatment failure, poor prognosis, poor specificity, etc.

Pending Publication Date: 2021-10-12
SHENZHEN LUOHU PEOPLELS HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide the application of CD137 antibody in the preparation of drugs that promote NK cells to express CD16 molecules and / or promote NK cells to secrete cytotoxic factors, aiming to solve the problems existing in existing tumor therapy drugs, especially breast cancer therapy drugs. poor specificity, leading to treatment failure and poor prognosis

Method used

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  • Application of CD137 antibody in preparation of drugs for promoting NK cells to express CD16 molecules
  • Application of CD137 antibody in preparation of drugs for promoting NK cells to express CD16 molecules
  • Application of CD137 antibody in preparation of drugs for promoting NK cells to express CD16 molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Culture of NK cells: Use human lymphocyte separation medium to separate peripheral blood mononuclear cells (PBMC) from heparin sodium anticoagulated peripheral blood derived from healthy volunteers, and follow the NK cell treatment kit and NK cell Instructions for serum-free medium for culturing NK cells. Follow up with NK cell serum-free medium for rehydration according to the growth status of NK cells. On the 0th, 8th, 12th, and 16th day of culture, samples were taken and counted. The micrographs of NK cells on the 8th and 16th day of culture are as follows: figure 1 shown. After 16 days of induced culture, the number of cells increased from the initial 3×10 6 cells expanded to 1.2×10 9 cells, expanded about 400 times, and the cell growth curve is shown as figure 2 shown;

[0053] (2) Phenotype detection of NK cells: take 1×10 6 NK cells on the 16th day of induction culture were washed once with DPBS, stained with CD3-FITC, CD56-PE, CD16-APC or CD137-APC a...

Embodiment 2

[0056] (4) Culture of NK cells: Use human lymphocyte separation medium to separate peripheral blood mononuclear cells (PBMC) from heparin sodium anticoagulated peripheral blood derived from healthy volunteers, and follow the NK cell treatment kit and NK cell Instructions for serum-free medium for culturing NK cells. Follow up with NK cell serum-free medium for rehydration according to the growth status of NK cells. On the 0th, 8th, 12th, and 16th day of culture, samples were taken and counted. The micrographs of NK cells on the 8th and 16th day of culture are as follows: figure 1 shown. After 16 days of induced culture, the number of cells increased from the initial 3×10 6 cells expanded to 1.2×10 9 cells, expanded about 400 times, and the cell growth curve is shown as figure 2 shown;

[0057] (5) Phenotype detection of NK cells: take 1×10 6 NK cells on the 16th day of induction culture were washed once with DPBS, stained with CD3-FITC, CD56-PE, CD16-APC or CD137-APC a...

Embodiment 3

[0068] (7) Immunofluorescence staining: use EGFR antibody and green fluorescent secondary antibody to specifically label EGFR protein in breast cancer cells MDA-MB-453 and MDA-MB-231 to detect the expression level of EGFR protein; at the same time, use Phalloidin-Atto565 Mark actin in the cells, inoculate 5000 breast cancer cells MDA-MB-231 and MDA-MB-453 on cell slides, and continue to culture overnight in a 37°C incubator; discard the culture in the culture wells base, washed once with DPBS, added 4% formaldehyde solution, and incubated at room temperature for 20 minutes; discarded 4% formaldehyde solution, added DPBS to wash twice; added 0.1% PBST (PBS solution containing 0.1% Triton X-100) and treated at room temperature for 5 minutes, Wash 3 times with DPBS; add 5% BSA blocking solution, incubate at room temperature for 1 hour; discard the blocking solution, add 200 μL EGFR antibody diluent, and incubate overnight in a 4°C refrigerator; discard the primary antibody solutio...

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Abstract

The invention belongs to the technical field of medicines, and particularly relates to an application of a CD137 antibody in preparation of drugs for promoting NK cells to express CD16 molecules and / or promoting the NK cells to secrete cytotoxic factors. The CD137 antibody acts on the NK cells, so that the expression proportion of CD16 in the NK cells can be remarkably increased, the NK cells can be remarkably promoted to secrete more cytotoxic factors, and therefore the cell killing effect of the NK cells is promoted. Therefore, the CD137 antibody is used for preparing the drugs for promoting the NK cells to express CD16 molecules and / or promoting the NK cells to secrete cytotoxic factors; and the CD137 antibody is also used for preparing the drugs for promoting the cell killing effect of the NK cells.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to the application of CD137 antibody in the preparation of drugs for promoting NK cells to express CD16 molecules and / or to promote NK cells to secrete cytotoxic factors. Background technique [0002] Natural killer (NK) cells are the core component of human innate immunity and play an important role in the body's anti-tumor immunity. NK cells have the ability to distinguish between normal cells and tumor cells, and can directly kill tumor cells non-specifically. histocompatibility complex, MHC) unrestricted killing characteristics. The phenotype of NK cells is CD3 - CD56 + According to the expression of CD16 molecules on the surface of NK cells, NK cells can be further divided into CD16 neg , CD16 dim and CD16 bright Three subgroups. CD16 bright NK cells can effectively play a cytotoxic role and can produce a certain level of cytokines, while CD16 neg and CD1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P35/00
CPCC07K16/2878A61P35/00A61K2039/505
Inventor 刘韬陈雪梅
Owner SHENZHEN LUOHU PEOPLELS HOSPITAL
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