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Tumor vaccine and preparation method thereof

A tumor vaccine, tumor technology, applied in the field of tumor vaccine and its preparation

Pending Publication Date: 2019-04-16
BEIJING ZHONGTAI HENGJI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the application of CD44 molecules in the field of tumor immunotherapy

Method used

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  • Tumor vaccine and preparation method thereof
  • Tumor vaccine and preparation method thereof
  • Tumor vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] The preparation of this application tumor vaccine

[0071] 1. Experimental Materials and Reagents

[0072] The C57BL / 6 mice used in the experiment were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, weighing about 18g.

[0073] Fetal calf serum (FCS): purchased from PAN company;

[0074] RPMI-1640 medium: purchased from PAN company;

[0075] GM-CSF and IL-4 were purchased from Peprotech, USA;

[0076] 2. Experimental steps

[0077] (1) Isolating mouse femur bone marrow hematopoietic precursor cells;

[0078] (2) The isolated mouse bone marrow hematopoietic precursor cells were mixed with 2×10 6 / mL density cultured in RPMI-1640 medium supplemented with 50ng / mL GM-CSF, 50ng / mL IL-4, 10% fetal calf serum (FCS), temperature 37°C, CO 2 The concentration is 5v / v%, induced for 7 days, the cell pellet is collected, the sample is resuspended with normal saline, and the smear is observed under a microscope. The results are as...

Embodiment 2

[0083] The preparation of this application tumor vaccine

[0084] 1. Experimental materials and reagents

[0085] The C57BL / 6 mice used in the experiment were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, weighing about 18g.

[0086] Fetal calf serum (FCS): purchased from PAN company;

[0087] RPMI-1640 medium: purchased from PAN company;

[0088] GM-CSF and IL-4 were purchased from Peprotech, USA;

[0089] The anti-mouse CD44 antibody was purchased from Biolegend, USA, the catalog number is 156002.

[0090] 2. Experimental steps

[0091] (1) Isolating mouse femur bone marrow hematopoietic precursor cells;

[0092] (2) The isolated mouse bone marrow hematopoietic precursor cells were mixed with 2×10 6 / mL density cultured in RPMI-1640 medium supplemented with 50ng / mL GM-CSF, 50ng / mL IL-4, 10% FCS, temperature 37°C, CO 2 The concentration is 5v / v%, induced for 7 days, the cell pellet is collected, the sample is resuspended...

Embodiment 3

[0097] The tumor vaccine of the present application (that is, the dendritic cells (CD44) knocking out the CD44 gene - / - DC)) and conventional dendritic cell tumor vaccine (that is, mature dendritic cells (CD44 + / + DC)) comparison of the ability to activate the immune system of the recipient.

[0098] 1. Experimental materials and reagents

[0099] The C57BL / 6 mice used in the experiment were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, weighing about 18g.

[0100] Cytokines in serum were detected using Biolegend Cytokine Detection Kit (Cat. No. 740005).

[0101] anti-mouse H 2 K b ,Ia b , CD40, CD80, CD86, and CD44 fluorescent antibodies were purchased from Biolegend Company in the United States, and the article numbers were H 2 K b (114605), Ia b (116410), CD40 (124607), CD80 (104705), CD86 (105005), CD44 (156002).

[0102] The antibody used to label mouse peripheral blood CD8 cells was PE-labeled anti-CD8 antibody (a...

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Abstract

The invention provides a tumor vaccine and a preparation method thereof. The tumor vaccine comprises dendritic cells blocking CD44 molecular signal channels. The invention also provides the method forpreparing the tumor vaccine. The method comprises the steps that 1) single karyocytes are obtained from the peripheral blood or umbilical cord blood of a mammal to be induced into the mature dendritic cells, or medullary hematopoiesis precursor cells are obtained from the bone marrow of the mammal to be induced into the mature dendritic cells; 2) the CD44 molecular signal channels of the mature dendritic cells are blocked; 3) the mature dendritic cells blocking the CD44 molecular signal channels are collected. The tumor vaccine has good tumor resistance, a wide anti-tumor spectrum and good use safety.

Description

technical field [0001] The invention relates to a vaccine and a preparation method thereof, in particular to a tumor vaccine and a preparation method thereof. Background technique [0002] Dendritic cells are professional antigen-presenting cells in the immune system and play an important role in the initiation, activation, effector and homeostasis of immune cells. Since the discovery of dendritic cells, their application in the field of anti-tumor has become a hot spot in tumor immunity. With the help of the strong immune activation ability of dendritic cells, dendritic cell tumor vaccines exert tumor antigen-specific / non-specific activation effects on the immune system of recipients, reverse the suppressive immune microenvironment formed in pre-cancer or tumors, mediate Immune killing and elimination of precancerous mutant cells or tumor cells. [0003] In 2011, Provenge, as the first FDA-approved dendritic cell tumor vaccine, was launched for the treatment of prostate c...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00
CPCA61K9/0019A61K39/0011A61K2039/5154A61K2039/54A61P35/00
Inventor 李政媛王超
Owner BEIJING ZHONGTAI HENGJI BIOLOGICAL TECH CO LTD
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