摘要
目的观察 6A8α-甘露糖苷酶表达对人 B细胞株 3D5增殖反应的影响。方法以重组腺相关病毒颗粒为载体，将正义 6A8 DNA或反义 6A8 DNA转导入 EB病毒转化的人 B细胞株 3D5，用单抗 6A8a染色反应和 Con A结合试验确认 6A8α-甘露糖苷酶高表达和低表达。以野生型细胞及转导空载载体的细胞作对照， MTT法检测细胞对 B细胞生长因子（ BCGF）、葡萄球菌 Cowen I（ SAC）或 IL- 6刺激的增殖反应。结果转导正义 6A8 DNA的细胞高表达 6A8α-甘露糖苷酶，转导反义 6A8 DNA的细胞低表达 6A8α-甘露糖苷酶。转导正义 6A8 DNA的 3D5细胞在 SAC诱导下的增殖反应明显增强（ P< 0.05），但转导反义 6A8 DNA细胞的增殖反应无变化。对低分子量 BCGF或 IL- 6诱导的刺激，转导正义 6A8或反义 6A8对细胞的增殖反应均无影响。结论 6A8α-甘露糖苷酶活性增高使 3D5细胞对 SAC刺激的增殖反应增强。 6A8α-甘露糖苷酶活性变化对低相对分子质量 BCGF或 IL- 6诱导的增殖反应无影响。
To study the effect of 6A8α- mannosidase expression on the proliferative response of human B cell 3D5. Methods Recombination of adeno- associated virus vector（ rAAV） mediated delivery of sense 6A8 DNA or antisense 6A8 DNA into 3D5 cells, and monoclonal antibody 6A8α staining and Con A binding assay for determination of the change of 6A8α- mannosidase expression, MTT assay for proliferation detection of 3D5 cells driven by Staphylococcus aureus crude cell suspension formalin- fixed (SAC)， lower molecular weight B cell growth factor (LMW- BCGF)， or rIL- 6.Results The expression of 6A8α- mannosidase was enhanced in sense 6A8- transducted cells and reduced in antisense 6A8- transducted cells. In comparison with the wild type and the mock- transducted cells, the proliferative response of the sense 6A8- transducted cells to SAC stimulation was enhanced (P< 0.05). However, transduction with antisense 6A8 did not affect the response. In addition, transduction with either sense or antisense 6A8 had no effect on proliferation of 3D5 cells induced by LMW- BCGF or IL- 6. Conclusion The proliferative response to SAC stimulation was enhanced in the 3D5 cells with enhanced expression of 6A8α- mannosidase. Either enhancement or reduction of 6A8α- mannosidase expression had no effect on proliferation induced by LMW- BCGF or IL- 6.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2000年第6期529-532,共4页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(39840008)资助
