Anti-tumor associated antigen WT1 specificity CTL and preparation method thereof

A tumor-associated antigen specific technology, applied in the field of anti-tumor-associated antigen WT1-specific CTL and its preparation, can solve the problem of poor anti-tumor efficacy of tumor immune cells, improve uptake efficiency and antigen presentation ability, enhance anti-tumor Tumor immunity, enhanced anti-tumor effect

Inactive Publication Date: 2016-06-29
深圳市中美康士生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 to wrap tumor cell antigen WT1, through greatly Improve the uptake efficiency and antigen presentation ability of DC cells to tumor antigens, enhance their anti-tumor effect, and 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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  • Anti-tumor associated antigen WT1 specificity CTL and preparation method thereof
  • Anti-tumor associated antigen WT1 specificity CTL and preparation method thereof
  • Anti-tumor associated antigen WT1 specificity CTL and preparation method thereof

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preparation example Construction

[0039] The invention provides a preparation method of anti-tumor-associated antigen WT1 specific CTL, comprising the following steps:

[0040] Step 1, preparing cationic liposome carrier targeting dendritic cell C-type lectin receptor coated with tumor-associated antigen WT1;

[0041] Step 2, using the cationic liposome carrier obtained in the step 1 to load the tumor-associated antigen WT1 on mature DC cells;

[0042] Step three, using the mature DC cells obtained in step two to induce WT1-specific T lymphocytes and central memory T lymphocytes.

[0043] The invention uses C-type lectin-type sugar-modified cationic liposome as an antigen carrier to encapsulate tumor-associated antigens 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 composed of ...

Embodiment 1

[0045] Example 1 Preparation method of anti-tumor-associated antigen WT1-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-associated WT1 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-associated 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 m...

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Abstract

The invention provides antigen-specificity T lymphocyte prepared from nano-liposome loaded tumor associated antigen WTI with a directional targeting function.According to the technology for preparing the tumor associated antigen-specificity T lymphocyte, a novel nano-liposome loaded tumor associated antigen is adopted, tumor correlation serves as a target of treatment, more effective specific cytotoxic T lymphocyte with the more effective targeting killing tumor correlation can be induced, and causes of tumor can be treated directly according to causes of tumorigenesis and relapse.Dendritic cells which are quickly mature within 72 hours are adopted, and a time window of optimal treatment of a tumor patient is greatly shortened.Meanwhile, the antigen-specificity T lymphocyte is amplified by means of engineering cells, and high-proportion Tcm with higher killing capacity can be generated.The method for preparing the tumor associated antigen WTI specificity T lymphocyte is simple and easy to implement, an immunity target spot aims at tumor correlation, antigenicity is strong, and stability is good.

Description

technical field [0001] The invention relates to the technical field of life sciences, in particular to an anti-tumor-associated antigen WT1 specific CTL and a preparation method thereof. Background technique [0002] Malignant tumors are the second leading cause of death threatening human life and health. According to statistics, in 2008, there were 12.7 million new cases of malignant tumors in the world and 7.6 million deaths. The occurrence, invasion and metastasis of malignant tumors are regulated by multiple levels and factors, and the imbalance between oncogenes and tumor suppressor genes is one of the key links. Therefore, finding new or further revealing the functions of existing oncogenes or tumor suppressor genes is of great significance for elucidating the mechanism of malignant tumor occurrence and evolution and developing potential therapeutic targets. Wilms' tumor gene 1 (Wilms' tumor gene 1, WT1) was first cloned and identified as a tumor suppressor gene in W...

Claims

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Application Information

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IPC IPC(8): C12N5/0783A61K39/00A61P35/00
CPCA61K39/0011C12N5/0638C12N2501/2302C12N2501/2312C12N2501/2318C12N2502/1121C12N2506/11
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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