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Controlled-release hepatoma cell vaccine depending on granulocyte-macrophage colony-stimulating factor (GM-CSF) wrapped by nanoparticles

A technology of GM-CSF and liver cancer cells, applied in the field of liver cancer cell vaccines, to achieve the effect of easy absorption

Inactive Publication Date: 2012-10-24
NANJING XINSAIERSI BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Aiming at the above problems, the present invention adopts chitosan-dependent GM-CSF nano slow-release particles combined with inactivated liver cancer cells to prepare a new type of liver cancer GVAX vaccine, which solves the safety, GM-CSF secretion stability and timeliness of the existing GVAX tumor vaccines And other issues

Method used

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  • Controlled-release hepatoma cell vaccine depending on granulocyte-macrophage colony-stimulating factor (GM-CSF) wrapped by nanoparticles
  • Controlled-release hepatoma cell vaccine depending on granulocyte-macrophage colony-stimulating factor (GM-CSF) wrapped by nanoparticles
  • Controlled-release hepatoma cell vaccine depending on granulocyte-macrophage colony-stimulating factor (GM-CSF) wrapped by nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the preparation of loading GM-CSF chitosan nanoparticles

[0030] Materials: Chitosan (Chitosan, deacetylation degree 75-85%, 448877), sodium tripolyphosphate (TPP, 238503) were purchased from SIGMA-ALDRICH company; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM -CSF) was purchased from PEPROTECH (315-03), Mouse GM-CSF Platinum ELISA was purchased from eBioscience ( BMS283 ).

[0031] Chitosan was dissolved in 0.8% volume fraction of acetic acid solution to prepare a chitosan solution with a mass concentration of 1 mg / mL, and rmGM-CSF was dissolved in ddH 2 O is configured into a solution with a mass concentration of 150ug / mL, sodium tripolyphosphate dissolved in ddH 2 O is configured as a solution with a mass concentration of 1.5 mg / ml.

[0032] Reaction process: 1.8ml 1mg / mL chitosan solution was added to 3.2ml ddH 2 0, and drop 1ml (400ul 150ug / mL rmGM-CSF+600ul 1.5mg / ml TPP) solution into the above solution at a rate of 1...

Embodiment 2

[0033] Example 2: Anti-tumor experiment of mice with sustained-release hepatocellular carcinoma vaccine relying on nanoparticle-encapsulated GM-CSF

[0034] 1) Animals and cell lines

[0035] 6-8-week-old male C57BL / (H-2b) mice were used as experimental mice, mouse liver cancer cell line Hepa1-6 was used as mouse liver cancer model and vaccine preparation; mouse melanoma cell line B78H1-GM- CSF was purchased from Eli Lilly and Company, and the expression of MHC class I molecules in this cell is missing, and GM-CSF can be efficiently secreted. Both cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C and 5% CO2.

[0036] 2) Vaccine construction

[0037] The sustained-release mouse hepatoma cell vaccine (hereinafter referred to as the new GVAX mouse vaccine) that relies on nanoparticle-encapsulated GM-CSF, the chitosan nanoparticles prepared in Example 1 and the inactivated mouse hepatocellular carcinoma cell line Hepa1 -6, the traditional GVAX mouse ...

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Abstract

A controlled-release hepatoma cell vaccine depending on granulocyte-macrophage colony-stimulating factor (GM-CSF) wrapped by nanoparticles is prepared from chitosan nanoparticles carrying GM-CSF and inactivated hepatoma cell. The method for preparing the vaccine comprises the steps of: stirring the mixed solution which consists of GM-CSF, sodium tripolyphosphate and chitosan solution for 2-3 hours, then collecting the reaction liquid, centrifuging utilizing glycerol at 8000-12000rpm for 10-15 minutes, carrying out 15-20KHz ultrasound resuspension using 0.9% sodium chloride for 30-45 seconds after finishing the centrifuging to obtain the uniform chitosan nanoparticles carrying GM-CSF, wherein the wrapping rate is 87.08+ / -2.32% and the particle size is 100+ / -23.68 nanometers; and mixing and suspending the chitosan nanoparticles carrying GM-CSF and the inactivated hepatoma cell to prepare the controlled-release hepatoma cell vaccine depending on GM-CSF wrapped by nanoparticles. The vaccine is simple to prepare, and has strong safety and high effectiveness.

Description

technical field [0001] The invention relates to the field of liver cancer cell vaccines, in particular to a slow-release liver cancer cell vaccine relying on nanoparticle-encapsulated GM-CSF. Background technique [0002] Tumor immunotherapy is an emerging tumor treatment method, which has a high status in the comprehensive treatment of tumors, and tumor vaccines are an important part of tumor immunotherapy. Tumor cell vaccines are the earliest and most widely used tumor vaccines, and their clinical application prospects and market potential are quite large. [0003] GVAX vaccine, an inactivated whole tumor cell vaccine that can secrete GM-CSF, is currently a widely used tumor cell vaccine. Granulocyte-macrophage colony stimulating factor (GM-CSF) is a cytokine that strongly stimulates the proliferation, differentiation and maturation of antigen-presenting cells such as macrophages and dendritic cells ( figure 1 ). In the early 1990s, Dranoff and Jaffee compared the diffe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K47/36A61P35/00
Inventor 孙倍成陈晨侯嘉杰王学浩
Owner NANJING XINSAIERSI BIOLOGICAL TECH
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