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Antitumor stem cell antigen OCT4 (octamer-binding transcription factor 4) specific CTL (cytotoxic T lymphocyte) and preparation method thereof

A stem cell antigen and tumor stem cell technology, which is applied in the field of anti-tumor stem cell antigen OCT4-specific CTL and its preparation, can solve the problems of poor anti-tumor efficacy of tumor immune cells, improve uptake efficiency and antigen presentation ability, and enhance anti-tumor The effect of enhancing anti-tumor immunity

Inactive Publication Date: 2016-05-25
深圳市中美康士生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Aiming at the problem of poor anti-tumor efficacy of existing tumor immune cells, the present invention proposes to use cationic liposomes targeting C-type lectin receptors on dendritic cells as an antigen carrier, and wrap tumor stem cell antigen Oct4, through greatly Improve the uptake efficiency and antigen presentation ability of DC cells to tumor antigens, enhance their anti-tumor effect, combine engineered cells and cytokines to massively expand antigen-specific T cells and Tcm, thereby enhancing anti-tumor immunity

Method used

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  • Antitumor stem cell antigen OCT4 (octamer-binding transcription factor 4) specific CTL (cytotoxic T lymphocyte) and preparation method thereof
  • Antitumor stem cell antigen OCT4 (octamer-binding transcription factor 4) specific CTL (cytotoxic T lymphocyte) and preparation method thereof
  • Antitumor stem cell antigen OCT4 (octamer-binding transcription factor 4) specific CTL (cytotoxic T lymphocyte) and preparation method thereof

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preparation example Construction

[0039] The invention provides a method for preparing anti-tumor stem cell antigen OCT4-specific CTL, comprising the following steps:

[0040] Step 1, preparing cationic liposome carrier targeting dendritic cell C-type lectin receptor coated with tumor stem cell antigen OCT4;

[0041] Step 2, using the cationic liposome carrier obtained in the step 1 to load the tumor stem cell antigen OCT4 on mature DC cells;

[0042] Step three, using the mature DC cells obtained in step two to induce OCT4-specific T lymphocytes and central memory T lymphocytes.

[0043] The invention adopts C-type lectin-type sugar-modified cationic liposome as an antigen carrier, wraps tumor stem cell antigen, and is used for the preparation of antigen-specific T lymphocytes. The cationic liposome includes phosphatidylethanolamine bilayer, polyethylene glycol derivatized phosphatidylethanolamine and mannose; the C-type lectin type sugar is mannose or mannoside. The phospholipid bimolecular ball is compose...

Embodiment 1

[0045] Example 1 Preparation method of anti-tumor stem cell antigen OCT4-specific CTL

[0046] The preparation method of the present invention mainly includes three major steps, and this embodiment is a general preparation method of the present invention.

[0047] First, the sugar-modified cationic liposome antigen carrier of C-type lectin type is produced, and the specific operation is as follows:

[0048] Firstly, mannose or mannoside modified polyethylene glycol derivatized phosphatidylethanolamine phospholipids are prepared. Mannose or mannoside is connected to the amino group of polyethylene glycol derivatized phosphatidylethanolamine phospholipid through aldehyde-amino or hydroxyl-amino condensation, so as to obtain the polyethylene glycol derivatized phosphatidyl alcohol of mannose or mannoside ethanolamine phospholipids. Then the cationic lipid DOTAP and the polyethylene glycol derivatized phosphatidylethanolamine phospholipid of mannose or mannoside are respectively...

Embodiment 2

[0055] Example 2 Identification of mature phenotype of DC

[0056] In this embodiment, mononuclear cells are collected and separated from peripheral blood at a rate of 3.0-5.0*10 6 Cells / ml were suspended in AIM-V serum-free medium, and cytokines IL-4 and GM-CSF were added to the medium to induce DC formation, and then placed at 37°C, 5% CO 2Culture in an incubator. On the next day, add cationic liposomes wrapped with tumor stem cell OCT4 antigen to the DC cell culture medium at a concentration of 1-2.5ug / ml and incubate for 6 hours, and then add DC promoting cells to the DC cells. The maturation medium is further cultured for 24-48 hours to obtain DC cells loaded with tumor stem cell antigens. The mature phenotype identification results of DC are attached figure 1 . Figures A and B are the detection data of mDC maturation of two donors, CD83 is a marker of DC maturation, and CD80 and CD86 are co-stimulatory molecules. The simultaneous double-positive expression rate of mD...

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Abstract

The invention relates to an antitumor stem cell antigen OCT4 (octamer-binding transcription factor 4) specific CTL (cytotoxic T lymphocyte) and a preparation method thereof. A directionally-targeted nano-liposome-carried tumor stem cell antigen is adopted to prepare the antigen-specific T lymphocyte. According to the tumor stem cell antigen-specific T lymphocyte preparation technique, the novel nano-liposome-carried tumor stem cell antigen is adopted, and the tumor stem cell is used as the therapeutical target, thereby inducing the more efficient targeted killing on the specific CTL of the tumor stem cell, and directly aiming at the tumor generation and relapse roots; and thus, the tumor therapy can implement a permanent cure. The 72-hour quick-maturated dendritic cell is adopted to greatly shorten the optimal therapeutical time window for the tumor patient. The engineering cell is adopted to amplify the antigen-specific T lymphocyte, thereby generating a high proportion of Tcm with higher killing capacity. The preparation method of the tumor stem cell antigen OCT4 specific T lymphocyte is simple and easy to implement; the immune target spot aims at the tumor stem cell; and thus, the antitumor stem cell antigen OCT4 specific CTL has high antigenicity and favorable stability.

Description

technical field [0001] The invention relates to the technical field of life sciences, in particular to an anti-tumor stem cell antigen OCT4-specific CTL and a preparation method thereof. Background technique [0002] In my country, malignant tumors have become one of the most important diseases that endanger human health, with an annual incidence rate of 235.23 / 100,000 and a mortality rate of 148.81 / 100,000. In recent years, with the development and improvement of technology, medical diagnosis and treatment, the overall mortality and survival rate of tumors have not improved significantly, and recurrence and metastasis are still the main reasons for the poor prognosis of cancer. With the in-depth study of the biological characteristics of tumors, some malignant solid tumors (glioma, lung cancer, breast cancer, and colorectal cancer, etc.) tumor initiating cells. Such cells only account for a small part of the whole tumor cells, but they have the ability of self-renewal and...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/87A61K39/00A61P35/00
CPCC07K14/47A61K39/00C12N5/0638C12N2501/2302C12N2501/2312C12N2501/2318C12N2506/11
Inventor 陈尚刘根桃李晓祥
Owner 深圳市中美康士生物科技有限公司
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