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Culture method of DC cell for enhancing CTL immune response

A cell culture, immune response technology, applied in the direction of blood/immune system cells, animal cells, vertebrate cells, etc., can solve the problems of ineffective activation of cytotoxic T cell proliferation and killing ability, long cycle, etc., to achieve effective in vivo. The effect of killing tumor, promoting proliferation and improving proliferation

Active Publication Date: 2015-11-04
丛秀丽
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Problems solved by technology

[0015] The technical problem to be solved by the present invention is to overcome the defects of the long cycle of DC obtained in the prior art, the ability to ingest and present tumor antigens is only 15-20%, and the proliferation and killing ability of cytotoxic T cells (CTL) cannot be effectively activated , providing an effective DC culture method is a way to improve the efficacy of immunotherapy

Method used

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  • Culture method of DC cell for enhancing CTL immune response
  • Culture method of DC cell for enhancing CTL immune response
  • Culture method of DC cell for enhancing CTL immune response

Examples

Experimental program
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Embodiment

[0065] A DC cell culture method for enhancing CTL immune response, comprising the following steps:

[0066] 1), collect blood, prepare adherent mononuclear cells;

[0067] ①. Collect freshly collected peripheral blood or umbilical cord blood in a blood collection bag containing heparin sodium anticoagulant, shake it up quickly to avoid coagulation; when it needs to be transported, put the blood bag and biological ice bag together in a special incubator Keep the temperature at 4-25°C, and the temperature difference should not exceed ±5°C. Ships to the lab within 6 hours. During transportation, ensure that the blood is not irradiated by X-rays and kept away from radiation sources.

[0068] ②. Dilute and mix the blood with an equal volume of normal saline. According to the volume ratio of the diluted blood to the lymphocyte separation medium is 3:2, slowly superimpose the diluted blood on the lymphocyte separation medium in the centrifuge tube. Pay attention to keep the liquid...

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Abstract

The invention discloses a culture method of DC cell for enhancing CTL immune response. According to the method, expression of Ube1L in the protein ubiquitination pathway is knocked out by sh-RNA, and tumor antigen-presenting ability of DC cell is effectively enhanced. More effective in-vivo tumor-killing capacity is exerted. A clinical-grade serum-free medium is used for culture of DC cell. By adding 1% human albumin and changing dose proportion of cytokines, culture time is shortened to 4 days, and time, space and personnel occupancy are minimized. By the method, types and dosage of a maturity-accelerating cytokine combination are adjusted, and the mature DC cell obtained can more effectively promote CD8+CTL cell proliferation. Therefore, antineoplastic curative effect is enhanced.

Description

technical field [0001] The invention relates to the field of cell preparation, in particular to a DC cell culture method for enhancing CTL immune response. Background technique [0002] CTL: also known as cytotoxic T cells (cytotoxic lymphocyte, CTL): a subset of white blood cells, a specific CD8+ T cells, specialized in secreting various cytokines to participate in immune function. It has a killing effect on antigenic substances such as certain viruses and tumor cells, and forms an important line of defense for the body's anti-virus and anti-tumor immunity with natural killer cells. The core of an effective anti-tumor immune response is to generate a cellular immune response with CD8+ T cells as the main body, which is also the basis of DC as an immunotherapy method. [0003] Protein ubiquitination modification: it is the main step in the degradation of intracellular proteins by the ubiquitin-proteasome system. A series of enzymatic activities such as ubiquitin ligase (ub...

Claims

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Application Information

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IPC IPC(8): C12N5/0784
Inventor 丛秀丽
Owner 丛秀丽
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