小鼠骨髓DC 2-DE技术体系建立及其蛋白质表达分析
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摘要
树突状细胞是机体中功能最强的专职抗原递呈细胞,既能直接激活初始型T淋巴细胞,启动早期特异性免疫应答;又能诱导免疫耐受,具有强大的激活CTL及CD4+ Th细胞的能力,控制着体内免疫反应的过程,在免疫应答中处于中心位置,已成为免疫学及相关学科的研究热点。
髓系树突状细胞分化发育的各阶段具有不同的功能特点。未成熟期树突状细胞可由粘附分子选择素介导进入炎症组织,并具有极强的抗原内吞和加工处理能力,但其激活初始T细胞的能力较弱。利用DC来增强机体特异性抗肿瘤免疫能力的研究,已成为肿瘤生物治疗的重要课题。同时,国内外都已将其作为肿瘤疫苗应用于基因治疗,取得了很好的临床效果。如果将抗原引入未成熟树突状细胞,使之发育为成熟细胞,则在临床应用上更为理想。
仙台病毒载体是细胞质型RNA载体,对各种动物细胞有较高的感染效率;在理论上不存在改变细胞核内染色体的危险性,与以往的载体相比具有更高的安全性。仙台病毒载体作为基因治疗、基因疫苗、基因(蛋白质)功能分析及蛋白质生产研究的载体,受到许多学者的青睐。
本课题对小鼠骨髓来源的未成熟树突状细胞的蛋白质表达进行了分析,通过双向电泳和MALDI-TOF MS分析,运用MS-Fit工具进行鉴定;对仙台病毒载体处理的成熟DC进行比较蛋白质组分析,寻找
Being the most potent professional antigen presenting cells, the Dendritic cells (DC) is endowed with the unique ability to stimulate naive T cells to initiate primary immune responses. In addition, the DC plays a critical role in the maintenance of self-tolerance by curtailing T cell responses directly or indirectly through the generation of CTL and CD4+Th regulatory cells. Many researchers have made emphasis on it in immunology and other related course.
The myeloid DC has different function during their different differentiation and development stage. Mediated by adhesion molecule, the DC enters inflammation tissue with the potent ability of antigen endocytosis and treatment, but it is endowed with the weak ability to stimulate naive T cells. It is an important topic for tumor biology therapy to enhance the body special immune ability to anti cancer by researched on DC. And it has afforded good clinical results that the DC is made as tumor vaccines for gene therapy at home and abroad. If we make the immature DC into mature DC by introducing antigens, the vaccines will be better at clinical application.
The negative-strand RNA viral vector system based on the Sendai virus (SeV) is able to efficiently infect many animal cells and more safer than other vectors because it can not change the chromosomes in the nucleus in theory. People like it as the vector for gene therapy, gene vaccine, analysis of gene function and protein production.
髓系树突状细胞分化发育的各阶段具有不同的功能特点。未成熟期树突状细胞可由粘附分子选择素介导进入炎症组织,并具有极强的抗原内吞和加工处理能力,但其激活初始T细胞的能力较弱。利用DC来增强机体特异性抗肿瘤免疫能力的研究,已成为肿瘤生物治疗的重要课题。同时,国内外都已将其作为肿瘤疫苗应用于基因治疗,取得了很好的临床效果。如果将抗原引入未成熟树突状细胞,使之发育为成熟细胞,则在临床应用上更为理想。
仙台病毒载体是细胞质型RNA载体,对各种动物细胞有较高的感染效率;在理论上不存在改变细胞核内染色体的危险性,与以往的载体相比具有更高的安全性。仙台病毒载体作为基因治疗、基因疫苗、基因(蛋白质)功能分析及蛋白质生产研究的载体,受到许多学者的青睐。
本课题对小鼠骨髓来源的未成熟树突状细胞的蛋白质表达进行了分析,通过双向电泳和MALDI-TOF MS分析,运用MS-Fit工具进行鉴定;对仙台病毒载体处理的成熟DC进行比较蛋白质组分析,寻找
Being the most potent professional antigen presenting cells, the Dendritic cells (DC) is endowed with the unique ability to stimulate naive T cells to initiate primary immune responses. In addition, the DC plays a critical role in the maintenance of self-tolerance by curtailing T cell responses directly or indirectly through the generation of CTL and CD4+Th regulatory cells. Many researchers have made emphasis on it in immunology and other related course.
The myeloid DC has different function during their different differentiation and development stage. Mediated by adhesion molecule, the DC enters inflammation tissue with the potent ability of antigen endocytosis and treatment, but it is endowed with the weak ability to stimulate naive T cells. It is an important topic for tumor biology therapy to enhance the body special immune ability to anti cancer by researched on DC. And it has afforded good clinical results that the DC is made as tumor vaccines for gene therapy at home and abroad. If we make the immature DC into mature DC by introducing antigens, the vaccines will be better at clinical application.
The negative-strand RNA viral vector system based on the Sendai virus (SeV) is able to efficiently infect many animal cells and more safer than other vectors because it can not change the chromosomes in the nucleus in theory. People like it as the vector for gene therapy, gene vaccine, analysis of gene function and protein production.
引文
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