基于比较转录组学的七鳃鳗类T、B淋巴细胞免疫应答信号传导分子机制

项目名称： 基于比较转录组学的七鳃鳗类T、B淋巴细胞免疫应答信号传导分子机制

项目编号： No.31271323

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 刘欣

作者单位： 辽宁师范大学

项目金额： 75万元

中文摘要： 本研究以无颌类脊椎动物日本七鳃鳗为对象，以生物信息学、比较基因组学、分子生物学及免疫学的研究方法,在对其成体造血组织脊椎弓脂肪体免疫前后各转录组之间及与高等脊椎动物的转录组进行比较分析的基础上，甄选在类T、B淋巴细胞活化、增殖过程中参与信号传导的其所特有的及与脊椎动物同源的信号传导分子。利用基因芯片技术检验差异表达候选基因并预测信号传导调控网络。筛选关键节点分子进行基因克隆，并进行重组蛋白表达、纯化及制备单克隆抗体；用流式细胞术根据分子标识筛选类淋巴细胞各亚群进行细胞学检验，用免疫荧光显微术检验这些分子在各类淋巴细胞亚群中的单一和共定位；采用免疫共沉淀技术研究这些分子的相互作用模式；验证并确立免疫激发后各类群淋巴细胞活化增殖过程的信号传导调控网络。阐明无颌类脊椎动物适应性免疫细胞各亚群免疫应答的分子遗传基础及作用机理。为揭示脊椎动物适应性免疫系统淋巴细胞信号转导体系的起源与进化提供新证据。

中文关键词： 七鳃鳗；类淋巴细胞；免疫应答；比较转录组；信号转导网络

英文摘要： In this study, lamprey (Lampetra japonicum), a jawless vertebrate, is targeted to investigate its signal transduction molecules of its own or homolougues to higher vertebrates involved in the activation and proliferation processes of T and B lymphocyte-like cells by means of bioinformatics, biochemistry, molecular biology and immunology research methods. These signaling molecules were mined, screened and identified from high throughput sequencing data of transcriptomes of dorsal fat body, a hematogenous tissue, before and after immune stimulation. The identified candidates are further verified by using gene chip technology, and gene regulatory signal transduction networks are predicted by bioinformatics methods. The key node elements in signal transduction networks are selected for gene cloning, recombinant protein expression, purification and preparation of monoclonal antibodies. These monoclonal antibodies of the key molecules are used to screen and verify corresponding class of lymphocyte subsets by flow cytometry and submicroscopic method. The localization and co-localization of these key molecules in various lymphocyte subsets are identified by Confocal Laser Scanning Microscope. The interaction modes of these molecular are investigated using immunoprecipitation techniques. The signal transduction regulator

英文关键词： Lamprey；lymphocyte-like cells；immune response；comparative transcriptome；transduction networks