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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
OBJECTIVE: To investigate the effect of transiently expressed antisense CD4 ORNA on the expression of CD4 0 and the proliferation of B cell membrane in healthy human being transformed by Epstein-Barr virus. Methods: The eukaryotic expression vector pcDNA3 / CD4 0 of human antisense CD4 O RNA was constructed by T A cloning and subcloning techniques and transfected into EB virus-transformed healthy human B cells. Flow cytometry (FACS) was used to detect the change of CD4 molecule expression on B cell membrane. The effect of antisense CD4 ORNA on proliferation of B cells was detected by MTT assay. Results: The expression of CD4 (superscript +) on transfected pcDNA3 / CD4 0 cells was significantly decreased (P <0.01) compared with the pcDNA3 group transfected with empty vector. The proliferation ability of pcDNA3 / CD4 0 was significantly decreased (P <0.01). Conclusion: Antisense CD4 0RNA technology can be used as an effective immunomodulatory tool. The CD4 0 gene itself plays an important role in cell growth and metabolism