反义CD40RNA对EB病毒转化的B细胞CD40分子表达和增殖能力的影响

目的 :探讨瞬时表达的反义CD4 0RNA ,对EB病毒转化的健康人B细胞膜表面CD4 0分子表达和增殖能力的影响。方法 :应用T A克隆技术和亚克隆技术 ,构建人反义CD4 0RNA的真核表达载体pcDNA3/CD4 0 ,并以其转染本室建立的EB病毒转化的健康人B细胞。应用流式细胞仪 (FACS) ,检测B细胞膜上CD4 0分子表达的变化。应用MTT比色法检测反义CD4 0RNA对B细胞增殖能力的影响。结果 :与转染空载体pcDNA3组相比 ,转染pcDNA3/CD4 0细胞上CD4 0分子的表达降低 (P<0 .0 1) ,其增殖能力明显降低 (P <0 .0 1)。结论 :反义CD4 0RNA技术 ,可作为有效的免疫调控手段。CD4 0基因本身在细胞的生长代谢中也起着重要作用

Effect of antisense CD40 RNA on CD40 expression and proliferation of EB virus-transformed B lymphocytes

AIM: To investigate the changes of CD40 expression and proliferation pattern of EB virus transformed B lymphocytes that had been transfected by antisense CD40 RNA. METHODS: Eukaryotic expression vector pcDNA3/CD40 of human CD40 antisense RNA was constructed by means of T A cloning and subcloning, and then was transfected into EB virus transformed B lymphocytes. Changes of CD40 expression on the B cells and their proliferation were tested by flow cytometry and MTT colorimetry assay, respectively. RESULTS: CD40 expression and proliferation of the B cells transfected with antisense CD40 RNA were significantly lower than those of the cells transfected with pcDNA3 alone( P <0.01). CONCLUSION: Antisense CD40 RNA may be served as an effective means of regulating immune function, and the CD40 gene itself also play an important role in cell proliferation.