High-yield T lymphocyte cloning technology for finding tumor specific antigen and tumor specific antigen determinant

A tumor-specific, lymphocyte-based technology, applied in the field of high-yield T-lymphocyte cloning technology, can solve problems such as T-lymphocyte death

Inactive Publication Date: 2005-04-27
SHANGHAI PUFAN BIOLOGICAL SCI TECH
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  • Abstract
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Problems solved by technology

In addition, the commonly used reverse immunization method obtains T lymphocytes by co-culturing in vitro with peripheral blood mononuclear cells pulsed with peptides, which also transmits signals through the TCR / CD3 pathway. As a result, the existing T lymphocytes relying on OKT-3 Cell expansion and cloning techniques likely lead to active guided death of tumor-specific resistant T lymphocytes
[0013] 2. Due to the technical limitations of current limiting dilution methods, there is no effective high-throughput method to discover tumor-specific antigens on a large scale so far

Method used

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  • High-yield T lymphocyte cloning technology for finding tumor specific antigen and tumor specific antigen determinant

Examples

Experimental program
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Embodiment 1

[0043] As shown in Figure 1, a typical example of HITC technology can be described as follows:

[0044] 1. Identify and expand tumor-reactive T lymphocytes

[0045] 1. Excision of tumor samples: Tumor samples are obtained from patients.

[0046] 2. Single-cell suspension: A part of the tumor sample is treated with various enzymes or processed by automatic mechanical force to make it into a single-cell suspension.

[0047] 3. Cell separation: use antibody-coated magnetic beads to separate CD4-positive and CD8-positive cells, while the remaining cells (mainly tumor cells) are cultured as attached cells.

[0048] 4. Limiting dilution with STIMULEUKIN (new auxin mixture): Based on the need for T lymphocyte cloning, the isolated CD4-positive and CD8-positive T lymphocytes are diluted, and then point to cultures that have been killed by irradiation. Cell-covered 96-well plates, with a maximum of one cell per well, are injected with STIMULEUKIN to expand these single-cell clones. ...

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Abstract

The present invention discloses the method of finding antigen determinant with tumor specificity, shown by HLA acceptor and identified by CD4 positive lymphocyte or CD8 positive lymph cell from the tumor sample and tumor penetrating lymphocyte gathered from patient body. The present invention discloses also the method of screening the latent curative effect of treating vaccine or immunogen in cell immunizing therapy on patient of specific race. The present invention discloses also the method of extracorporeal amplifying to generate T-lymphocyte with specific reaction on antigen and antigen determinant for cell transferring therapy or developing treating vaccine to activate cell immunity system.

Description

technical field [0001] The invention relates to the field of tumor immunology, and further, the invention relates to a high-yield T lymphocyte cloning technique for discovering tumor-specific antigens and tumor-specific antigenic determinants. Background technique [0002] Many studies have shown that the control and treatment of cancer can be achieved by manipulating the human immune system. For example, using the body's mucus immune response, monoclonal antibodies can be developed against various growth factor receptors and cell surface marker molecules to achieve the goal of treating cancer. effect; on the other hand, the use of cellular immune response to activate the human immune system to specifically kill tumor cells to achieve the treatment of cancer has been paid more and more attention. [0003] The cellular immune response of the human immune system is mainly realized through T lymphocytes (Tlymphocytes). T lymphocytes can be divided into CD8-positive killer T ly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K45/00A61P37/04C12N5/0783C12P19/34C12P21/00G01N33/53
Inventor 范青青曾钢
Owner SHANGHAI PUFAN BIOLOGICAL SCI TECH
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