Ding Fei-Xiang, Xian Xian, Guo Ying-Jun, Liu Yin, Wang Yue, Yang Fu, Wang Yu-Zhao, Song Shu-Xia, Wang Fang, Sun Shu-Han
Department of Medical Genetics, The Second Military Medical University, Shanghai 200433, P R China.
Mol Biosyst. 2010 Nov;6(11):2192-9. doi: 10.1039/c005222a. Epub 2010 Aug 27.
Hepatitis B virus core protein virus-like particles (HBc-VLPs) act as a strong immunogen and are suitable for uptake by dendritic cells (DCs), in which they directly promote DC maturation and migration. To illustrate the utility of global proteomic analysis techniques in elucidating the molecular events that are altered in HBc-VLP-pulsed bone marrow-derived DCs (BMDCs) and to gain a better understanding of the molecular mechanisms of capture and processing of HBc-VLP-pulsed BMDCs, an antigen (Ag) delivery system based on HBc-VLP-pulsed BMDCs was developed. Two-dimensional electrophoresis (2-DE) and tandem mass spectrometry (MS/MS) analyses were utilized to analyze the differential protein expression patterns between HBc-VLP-pulsed and untreated BMDCs. Protein spots with significantly altered expression levels were detected, identified and validated. The results showed that exogenous HBc-VLPs were phagocytosed efficiently by BMDCs and enhanced the efficacy of BMDC maturation and Ag presentation, VLPs also induced high levels of Ag-specific CD8(+) T cells that displayed high cytotoxic T lymphocyte (CTL) activity in vivo. Several differentially expressed proteins, including growth factor receptor bound protein 2 (Grb2) and annexin A2 (AnxA2), were detected by proteomic analysis, identified by mass spectrometry and validated by western blot.
乙肝病毒核心蛋白病毒样颗粒(HBc-VLPs)是一种强大的免疫原,适合被树突状细胞(DCs)摄取,在DCs中,它们直接促进DC的成熟和迁移。为了阐明全球蛋白质组分析技术在揭示HBc-VLP脉冲骨髓来源的DCs(BMDCs)中发生改变的分子事件方面的效用,并更好地理解HBc-VLP脉冲BMDCs的捕获和加工分子机制,开发了一种基于HBc-VLP脉冲BMDCs的抗原(Ag)递送系统。利用二维电泳(2-DE)和串联质谱(MS/MS)分析来分析HBc-VLP脉冲和未处理的BMDCs之间的差异蛋白质表达模式。检测、鉴定并验证了表达水平有显著改变的蛋白质斑点。结果表明,外源性HBc-VLPs被BMDCs有效吞噬,增强了BMDC成熟和Ag呈递的功效,VLPs还诱导了高水平的Ag特异性CD8(+) T细胞,这些细胞在体内表现出高细胞毒性T淋巴细胞(CTL)活性。通过蛋白质组分析检测到几种差异表达的蛋白质,包括生长因子受体结合蛋白2(Grb2)和膜联蛋白A2(AnxA2),通过质谱鉴定并通过蛋白质印迹验证。