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B细胞的激活是通过B细胞受体与其特异性抗原的结合引起,并引发后续的信号级联反应.深入了解这一重要事件的分子机制是免疫学研究的重要目标.嵌合B细胞受体是B细胞信号功能研究的有效工具.然而,这一方法仅适用于工具细胞,不能被用于体内研究.本研究构建了能同时表达针对特异性抗原的膜型免疫球蛋白的重链和轻链的逆转录病毒载体,通过逆转录病毒载体将这一膜型免疫球蛋白表达在原始B淋巴细胞膜上.结果证实,通过逆转录病毒载体表达的膜型免疫球蛋白能引发B细胞受体的信号转导,可引发B细胞受体下游Syk和Erk1/2蛋白的磷酸化.本研究进一步表明,B淋巴细胞表达的膜型免疫球蛋白在体外和体内均能与特异性抗原结合并导致细胞增殖.因此,本研究提供了一个能在体外和体内快速方便地分析B细胞受体下游信号级联反应的方法,这一方法将有助于更深入地研究与B细胞功能相关的各种蛋白.
The activation of B cells is caused by the binding of B cell receptors to their specific antigens and triggers subsequent signaling cascades.It is an important goal of immunological studies to understand the molecular mechanism of this important event.Chimeric B cell receptor Is an effective tool for the study of the function of B cell signaling.However, this method is only applicable to tool cells and can not be used for in vivo studies.In this study, we constructed a recombinant plasmid that can simultaneously express the heavy and light immunoglobulin of the membrane-specific immunoglobulin Chain retroviral vector to express this membrane-type immunoglobulin on the original B lymphocyte membrane by means of a retrovirus vector It was confirmed that the membrane-type immunoglobulin expressed by the retrovirus vector can induce the B cell receptor Signal transduction can trigger the phosphorylation of Syk and Erk1 / 2 downstream of B cell receptor.The present study further shows that membrane-bound immunoglobulin expressed by B lymphocytes can bind with specific antigen in vitro and in vivo and cause cell Proliferation.Therefore, this study provides a rapid and convenient method for the analysis of signaling cascades downstream of B cell receptors in vitro and in vivo, which will help to further Study various proteins related to B cell function.