Application of beta-dextran in preparation of human dendritic cell tumor vaccine

A dendritic cell and tumor vaccine technology, which is applied in the application field of beta-glucan in the preparation of human dendritic cell tumor vaccine, can solve the problems such as undiscovered human dendritic cell tumor vaccine, and achieves the improvement of immunity. Potency, the effect of promoting proliferative ability

Active Publication Date: 2012-07-25
戚春建
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no literature report on the preparation of human dendritic cell tumor vaccine with β-glucan

Method used

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  • Application of beta-dextran in preparation of human dendritic cell tumor vaccine
  • Application of beta-dextran in preparation of human dendritic cell tumor vaccine
  • Application of beta-dextran in preparation of human dendritic cell tumor vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1）

[0021] The application of the β-glucan of this example in the preparation of human dendritic cell tumor vaccine has the following two steps:

[0022] ① Obtaining immature human dendritic cells:

[0023] Use the Fresenius blood cell separator and its matching disposable consumption tubing, select the double-needle mononuclear cell program, and input the corresponding parameters of the person to be collected (sex, height, weight, hematocrit, etc.), according to the collected person Adjust the blood flow rate to 50mL / min to 80mL / min according to the specific situation, and the ratio of anticoagulant to whole blood is 1:10. Mononuclear cells are separated from human peripheral blood, and the volume of processed whole blood does not exceed twice the volume of own blood. , to obtain peripheral blood mononuclear cells.

[0024] Place the peripheral blood mononuclear cells obtained above into a 15 mL centrifuge tube, take 10 μL of cells for counting, and then add anti-human CD14 + I...

experiment example 1

[0030] (Experimental example 1, immature human dendritic cells can phagocytose β-glucan)

[0031] Take 5×10 5 Step 1 of Example 1 ①The immature human dendritic cells obtained by culturing for 120 h and fluorescein DTAF-labeled β-glucan (concentration: 20 μg / mL) were cultured at 37°C for 1 h, immediately placed on ice, and washed with pre- Wash with cold PBS 3 times and perform flow cytometry. See the experimental results figure 2 .

[0032] Depend on figure 2 It can be seen that immature human dendritic cells can phagocytose β-glucan.

experiment example 2

[0033] (Experimental example 2, the effect of β-glucan on the expression of co-stimulatory molecules and the concentration of cytokines)

[0034] In this experimental example, three groups of human dendritic cells were used to detect CD83, CD86, CD40, HLA-DR, and CCR7 surface markers to determine maturity. The first group is the immature human dendritic cells collected by centrifugation in step ① of Example 1 and cultured for 120 hours. The second group was human dendritic cells 48 hours after the immature human dendritic cells obtained in the first group were stimulated by β-glucan (concentration: 100 μg / ml). The third group is the human dendritic cells obtained after stimulating the immature human dendritic cells obtained in the first group by TNF-α (concentration: 20ng / mL) for 48 hours. The experimental results are shown in image 3 .

[0035] Depend on image 3 It can be seen that the expression of co-stimulatory molecules in the second group significantly increased aft...

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Abstract

The invention discloses an application of beta-dextran in preparation of human dendritic cell tumor vaccine. The application comprises culturing human peripheral blood mononuclear cells with culture medium containing GM-CSF (granulocyte-macrophage colony-stimulating factor) and IL-4 (interleukin-4) to obtain immature human dendritic cell; and inducing loading of tumor antigen onto the immature human dendritic cell through beta-dextran with concentration of 80-120 mug / mL, to obtain human dendritic cell tumor vaccine. According to the invention, the human dendritic cell tumor vaccine is obtained by inducing loading of tumor antigen onto the immature human dendritic cell through beta-dextran. The vaccine can improve immunizing potency of human dendritic cell, promote multiplication capacity of T-cells, especially can externally stimulate the T-cells to achieve specific tumor killing ability, thereby treating various malignant solid tumors.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to the application of a beta-glucan in preparing a human dendritic cell tumor vaccine. Background technique [0002] Dendritic cells (DC) account for about 0.3% of the entire cell population in the blood. They are professional antigen-presenting cells that can stimulate and regulate the body's immune response. The following characteristics of dendritic cells enable it to effectively present antigens to stimulate the body's immune response: (1) It can efficiently capture antigens through phagocytosis, pinocytosis and receptor-mediated endocytosis (such as Apoptotic and necrotic cell microorganisms and soluble foreign proteins, etc.); (2) have strong motility and can transport captured antigens from peripheral tissues to primary and secondary lymphoid organs; (3) can express at a high level MHC-I and MHC-II molecules form complexes with antigenic determinants formed by in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/385A61K39/00C12N5/0784A61P35/00
Inventor 戚春建宁永玲王仕忠钱科卿
Owner 戚春建
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