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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: 2014-04-09
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] Example 1: Preparation of GM-CSF-loaded chitosan nanoparticles

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

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

[0032] Reaction process: 1.8ml 1mg / mL chitosan solution is added to 3.2ml ddH 2 0, and drop 1ml (400ul 150μg / mL rmGM-CSF+600ul 1.5mg / ml TPP) solution into the above solution at a speed of 100ul / min, continue ...

Embodiment 2

[0033] Example 2: Anti-tumor experiment of slow-release hepatoma cell vaccine relying on nano-particles encapsulating GM-CSF on mice

[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. This cell lacks expression of MHC class I molecules and can efficiently secrete GM-CSF. 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 hepatocarcinoma vaccine (hereinafter referred to as the new GVAX mouse vaccine) that relies on nanoparticles to encapsulate GM-CSF, the chitosan nanoparticles prepared in Example 1 and the inactivated mouse hepatocellular carcinoma cell line Hepa1 -6, the traditional GVAX mouse vaccine is composed of inactivated Hepa1-6 and B78H1...

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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 present invention relates to the field of liver cancer cell vaccines, in particular to a slow-release liver cancer cell vaccine that relies on nanoparticles to encapsulate GM-CSF. Background technique [0002] Tumor immunotherapy is an emerging tumor treatment method, which has a high position in comprehensive tumor treatment, and tumor vaccine is an important part of tumor immunotherapy. Tumor cell vaccines are the earliest research and most widely used tumor vaccines, and their clinical application prospects and market potential are quite large. [0003] GVAX vaccine, that is, an inactivated whole tumor cell vaccine that secretes GM-CSF, is currently a more 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...

Claims

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Application Information

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Patent Type & Authority Patents(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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