Method for preparing human dendritic cell vaccine

A technology of dendritic cells and vaccines, applied in the field of cellular immunology, to achieve the effect of simple process, low cost, and easy large-scale production

Active Publication Date: 2014-02-26
SHENZHEN HORNETCORN BIOTECH
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Problems solved by technology

[0003] At present, there are many routine methods for DC expansion, such as classic GM-CSF and IL-4 to induce DC differentiation, and then use TNF-α or a combination of cytokines IL-1β, IL-6, TNF-α, PGE-2, etc. Or combined with polyinosinic acid (polycytidylic acid, poly-I: C), bacter

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  • Method for preparing human dendritic cell vaccine
  • Method for preparing human dendritic cell vaccine

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Embodiment 1

[0024] Example 1 Preparation of human dendritic cell vaccine and detection of maturity and IL-12 secretion

[0025] The first step: further separate and obtain mononuclear cells from the isolated PBMCs, including the following steps:

[0026] (1) 50 ml of peripheral venous blood was collected from the patient, and mononuclear cells were obtained by density gradient centrifugation of Ficoll-diatrizoate glucosamine. The specific steps are: 1500 rpm, centrifuge for 10 minutes, absorb the upper plasma layer, inactivate at 56°C for 30 minutes and centrifuge for later use, dilute the precipitated blood cells with normal saline, and divide human lymphocyte separation medium and diluted blood at a ratio of 1:2 The proportion of the mixture was added to the centrifuge tube, 2000 rpm, centrifuged for 20 minutes, carefully sucked the buffy coat layer, washed twice with normal saline, the speeds were 1600 rpm, 1300 rpm, and centrifuged for 7 minutes to obtain the peripheral Blood mononuc...

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Abstract

The invention belongs to the technical field of cellular immunology, and particularly relates to a method for preparing a human dendritic cell vaccine. The method comprises the following steps: (1) separating a monocyte from peripheral blood mononuclear cells by an adherent method; (2) inducing the monocyte to differentiate into DC by using a DC culture medium, and adding a tumor antigen on the third day; (3) on the fifth day, adding a combined type DC maturity promoting agent of IL-1beta, IL-6, TNF-alpha, IFN-gamma and Mtb-HAg; and (4) on the sixth-seventh day, detecting the maturity degree of the DC and the secretion amount of IL-12. The prepared DC is mainly applied in treatment of cancer patients or prevention of high-risk cancer groups. The method has the advantages that the prepared DC can secrete a large amount of IL-12, thereby prompting T cells to differentiate in the direction of Th1-type immune response; and moreover, the method has the superiorities of simple preparation technology, low cost, easy large-scale production and the like.

Description

technical field [0001] The invention belongs to the technical field of cellular immunity, in particular to a preparation method of a human dendritic cell vaccine. Background technique [0002] Dendritic cell (DC) is the most powerful antigen-presenting cell (APC) known so far, and its biggest feature is that it can stimulate naive T cells ( T cell) activation and proliferation are the enablers of specific immune responses. Therefore, DCs have broad application prospects in tumor immune cell therapy. However, the proportion of DC in human peripheral blood is very low, and the function of DC in tumor patients is mostly in a suppressed state. Therefore, how to obtain a sufficient number of DCs capable of inducing T cells to differentiate into cytotoxic T lymphocytes (cytotoxic T lymphocytes, CTL) in the direction of Th1 immune response has become the key to clinical application. [0003] At present, there are many routine methods for DC expansion, such as classic GM-CSF and...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00C12N5/0784
Inventor 马飞王宇环罗晓玲
Owner SHENZHEN HORNETCORN BIOTECH
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