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Tumor vaccine and preparation method thereof

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

Pending Publication Date: 2019-05-24
BEIJING ZHONGTAI HENGJI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There is no report on the simultaneous application of CD44 molecules and CD11b 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

[0072] The preparation of this application tumor vaccine

[0073] 1. Experimental Materials and Reagents

[0074] 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.

[0075] Fetal calf serum (FCS): purchased from PAN company;

[0076] RPMI-1640 medium: purchased from PAN company;

[0077] GM-CSF and IL-4 were purchased from Peprotech, USA;

[0078] A lentiviral vector for simultaneously knocking out CD44 and CD11b genes: purchased from Beijing Hopson Genetics Company.

[0079] 2. Experimental steps

[0080] (1) Isolating mouse femur bone marrow hematopoietic precursor cells;

[0081] (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...

Embodiment 2

[0086] The preparation of the application tumor vaccine

[0087] 1. Experimental materials and reagents

[0088] 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.

[0089] Fetal calf serum (FCS): purchased from PAN company;

[0090] RPMI-1640 medium: purchased from PAN company;

[0091] GM-CSF and IL-4 were purchased from Peprotech, USA;

[0092] Anti-mouse CD44 antibody and anti-mouse CD11b antibody were purchased from Biolegend, USA, the catalog number of anti-mouse CD44 antibody was 156002, and the catalog number of anti-mouse CD11b antibody was 101248.

[0093] 2. Experimental steps

[0094] (1) Isolating mouse femur bone marrow hematopoietic precursor cells;

[0095] (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, temperat...

Embodiment 3

[0100] The tumor vaccine of the present application (that is, dendritic cells (CD11b knockout CD44 and CD11b gene) - / - CD44 - / - DC)) and conventional dendritic cell tumor vaccine (that is, mature dendritic cells (CD11b + / + CD44 + / + DC)) comparison of the ability to activate the immune system of the recipient.

[0101] 1. Experimental materials and reagents

[0102] 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.

[0103] Cytokines in serum were detected using Biolegend Cytokine Detection Kit (Cat. No. 740005).

[0104] anti-mouse H 2 K b ,Ia b , CD40, CD80, CD86, CD44, and CD11b 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), CD11b (101248).

[0105] The antibody used to label mouse peripheral blood CD...

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Abstract

The invention provides a tumor vaccine and a preparation method thereof. The tumor vaccine comprises dendritic cells which block signal pathways of CD44 and CD11b molecules. The invention further provides a method for preparing the tumor vaccine. The method includes the following steps: (1) acquiring mononuclear cells from peripheral blood or umbilical cord blood of a mammal, inducing the mononuclear cells into mature dendritic cells or acquiring bone marrow hematopoietic precursor cells from bone marrow of the mammal, and inducing the mononuclear cells into mature dendritic cells; (2) blocking the signal pathways of the CD44 and CD11b molecules of the mature dendritic cells; and (3) collecting the dendritic cells blocking the signal pathways of the CD44 and CD11b molecules. The tumor vaccine has a good anti-tumor effect, 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): A61K39/00A61P35/00C12N5/0784
Inventor 李政媛王超
Owner BEIJING ZHONGTAI HENGJI BIOLOGICAL TECH CO LTD
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