Antigen activated immune cells and culturing method and application thereof

A culture method and technology of immune cells, which are applied to the field of antigen-activated immune cells and their culture, can solve the problems of long culture acquisition time, single cell type, exogenous infection, etc., and achieve short culture time, strong antigen specificity, The effect of avoiding uncertainty

Active Publication Date: 2016-08-31
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to overcome the above-mentioned deficiencies in the prior art, and provide an immune cell activated by antigen and a culture method thereof, so as to solve the problem of a single type of cultured cells in the existing DC cell culture; 2. The technical problem that the culture period is relatively long
[0012] Another object of the present invention is to provide an anti-tumor drug to overcome the risk of exogenous infection and long time for culture and acquisition in other existing cell therapy drugs, which delay the risk of tumor dissemination in patients

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  • Antigen activated immune cells and culturing method and application thereof
  • Antigen activated immune cells and culturing method and application thereof

Examples

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Embodiment 1

[0065] This embodiment provides a method for culturing antigen-activated immune cells and the antigen-activated immune cells cultured by the method. The antigen-activated immune cell culture method comprises the following steps:

[0066] S11. Peripheral blood mononuclear cell (PMBC) suspension preparation:

[0067] Take 50ml of peripheral blood from the patient, dilute it with normal saline 1:1, add 15ml of lymphocyte separation medium, and centrifuge (2000rpm, 20min) to collect buffy coat cells;

[0068] Wash three times with 50ml saline, and finally resuspend with 1640 culture medium (12ml) containing 100ng / ml GM-CSF and 10ng / ml IL-14;

[0069] Induced amplification of S12.AAIC:

[0070] The PMBC suspension prepared in step S11 was divided into six-well plates, and the PMBC concentration was 5×10 6 / ml, 2ml / well, add the corresponding antigen NY-ESO-1, 1000U / ml;

[0071]Placed in saturated humidity, 37°C, 5.0% CO 2 Cultivated in an incubator;

[0072] Harvest AAIC on t...

Embodiment 2

[0075] This embodiment provides a method for culturing antigen-activated immune cells and the antigen-activated immune cells cultured by the method. The steps of the antigen-activated immune cell culture method are as in Example 1, except that the tumor antigen is prostate cancer-specific antigenic protein (PSA), and the cytokines are IL-2, IL-14 and G-CSF. Combination, wherein the final concentration of PSA is 500u / ml, the final concentration of IL-2 is 1000u / ml, the final concentration of IL-14 is 20ng / ml, and the final concentration of G-CSF is 150ng / ml.

Embodiment 3

[0077] This embodiment provides a method for culturing antigen-activated immune cells and the antigen-activated immune cells cultured by the method. The steps of the antigen-activated immune cell culture method are the same as those in Example 1, except that the tumor antigen is a melanoma-specific antigen (MAGE-3), and the cytokines are IL-2, IL-14 and G-CSF The combination, wherein the final concentration of MAGE-3 is 1000U / ml, the final concentration of IL-2 is 500u / ml, the final concentration of IL-14 is 50ng / ml, and the final concentration of G-CSF is 100ng / ml.

[0078] The AAICs induced and amplified in Examples 2 and 3 were cultured in vitro, and the cultured products were subjected to gel electrophoresis test. It was known that the AAIC stimulated the body to secrete IL-2, TNF-α, IFN-γ and other Antitumor cytokines.

[0079] From the in vitro experiments of the AAIC provided in Examples 1-3, it is known that the AAIC provided in the embodiment of the present invention...

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Abstract

The invention discloses antigen activated immune cells and a culturing method and an application thereof. The culturing method of the antigen activated immune cells comprises the following steps: a step of collecting peripheral blood mononuclear cells of a patient, and a step of adding the mononuclear cells and tumor antigens and/or cytokines into a serum-free culture medium and carrying out activating culture. The antigen activated immune cells are used for preparation of drugs for cell treatment of tumor. The culturing method of the antigen activated immune cells adopts the serum-free culture medium in the whole process, and effectively avoids any possible exogenous infection; the antigen specificity is strong, multiple cells are activated, the role is closer to the whole effect of a human body anti-tumor role, and the culturing time is short. The antigen activated immune cells and the tumor cell-treatment drugs can produce a plurality of epitope specific anti-tumor effect networks in vivo through induction, and the anti-tumor role is jointly achieved through mutual collaboration of the effect networks.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an antigen-activated immune cell, a culture method thereof and an application thereof. Background technique [0002] DC cells, also known as dendritic cells, are a heterogeneous cell population with high-efficiency killing activity. The proportion of DC cells in peripheral blood lymphocytes is 1% to 5%. It has been clinically confirmed that after a large amount of DC expansion, It has significant tumor-killing and virus-clearing activities, and DC plays an important regulatory role in the body's cellular immunity and humoral immunity. [0003] DC cell immunotherapy is the most important part of biological therapy. The process is to extract the patient's own immune cells, activate, modify and proliferate them in vitro, and then infuse them back into the patient's body to induce the body to produce specific or non-specific immune responses. Play the role of killing tumor c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N5/0783C12N5/0784A61K39/00A61P35/00
CPCA61K39/0011A61K2039/5154A61K2039/5158C12N5/0634C12N5/0636C12N5/0639C12N2501/22C12N2501/2302C12N2501/2314C12N2501/998
Inventor 田耕夏燕毕冬梅李小燕肖奕丹
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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