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含MAGE-3肽纳米疫苗对小鼠移植性胃癌的制备及其抗肿瘤作用

Preparation and antitumor effects of nanovaccines with MAGE-3 peptides in transplanted gastric cancer in mice.

作者信息

Yang Jun, Li Zhi-Hua, Zhou Jia-Jia, Chen Ru-Fu, Cheng Liang-Zheng, Zhou Quan-Bo, Yang Li-Qun

机构信息

Department of Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, PR China.

出版信息

Chin J Cancer. 2010 Apr;29(4):359-64. doi: 10.5732/cjc.009.10541.

DOI:10.5732/cjc.009.10541
PMID:20346208
Abstract

BACKGROUND AND OBJECTIVE

As a prospective vaccine carrier, nanoparticles can protect antigens from degradation and enhance immune response. This study prepared nanovaccines with MAGE-3-derived CD4+-CD8+T cell epitope peptides, and investigated its character and antitumor effects on transplanted gastric cancer in mice.

METHODS

We adopted the self-assembly method to prepare peptide/chitosan conjugated with deoxycholic acid (chitosan-deoxycholic acid) nanoparticles. We observed the appearance of the chitosan-deoxycholic acidnanoparticles through a transmission electron microscope (TEM) and analyzed the peptide content and its release pattern by fluorescence spectrophotometry. We observed tumor-suppression efficacy in vivo through animal experiments.

RESULTS

We successfully prepared nanoparticles with MAGE-3 peptide antigen, and its encapsulation efficiency and loading level were about 37% and 17.0%, respectively. These nanoparticles presented a delayed release pattern in phosphate buffered saline (PBS) at pH 7.4, and the full release time was about 48 h. In 2 mg/mL lysozyme, the nanoparticles showed a sudden release, and the full release time was about 24 h. ELISPOT and cytotoxic experiments showed that the MAGE-3 peptide loaded nanoparticles could stimulate immune response in vivo and could generate MAGE-3-targeted cytotoxic T lymphocytes (CTLs), and kill MAGE-3-specific tumor cells. Tumor suppression experiments showed that the regression ratio of the peptide-loaded nanoparticles group was 37.81%.

CONCLUSIONS

MAGE-3 peptide/chitosan-deoxycholic acidvaccine-loaded nanoparticles can stimulate antitumor immune response in vivo and can regress the growth of mouse forestomach carcinoma cell line MFC.

摘要

背景与目的

作为一种潜在的疫苗载体,纳米颗粒可保护抗原不被降解并增强免疫反应。本研究制备了含有MAGE-3衍生的CD4⁺-CD8⁺T细胞表位肽的纳米疫苗,并研究其特性及对小鼠移植性胃癌的抗肿瘤作用。

方法

我们采用自组装方法制备了与脱氧胆酸偶联的肽/壳聚糖(壳聚糖-脱氧胆酸)纳米颗粒。通过透射电子显微镜(TEM)观察壳聚糖-脱氧胆酸纳米颗粒的外观,并通过荧光分光光度法分析肽含量及其释放模式。我们通过动物实验观察体内的肿瘤抑制效果。

结果

我们成功制备了含有MAGE-3肽抗原的纳米颗粒,其包封率和负载量分别约为37%和17.0%。这些纳米颗粒在pH 7.4的磷酸盐缓冲盐水(PBS)中呈现缓释模式,完全释放时间约为48小时。在2 mg/mL溶菌酶中,纳米颗粒呈现突释,完全释放时间约为24小时。ELISPOT和细胞毒性实验表明,负载MAGE-3肽的纳米颗粒可在体内刺激免疫反应,并可产生靶向MAGE-3的细胞毒性T淋巴细胞(CTL),并杀死MAGE-3特异性肿瘤细胞。肿瘤抑制实验表明,负载肽的纳米颗粒组的肿瘤消退率为37.81%。

结论

负载MAGE-3肽/壳聚糖-脱氧胆酸疫苗的纳米颗粒可在体内刺激抗肿瘤免疫反应,并可使小鼠前胃癌细胞系MFC的生长消退。

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