6A8α—甘露糖苷酶表达对人B淋巴细胞3D5增殖反应的影响

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。

Effect of 6A8 alpha-mannosidase expression on the proliferative response of human B cell line 3D5

OBJECTIVE: To study the effect of 6A8 alpha-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 6A8 alpha staining and Con A binding assay for determination of the change of 6A8 alpha-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 6A8 alpha-mannosidase was enhanced in sense 6A8-transduced cells and reduced in antisense 6A8-transduced cells. In comparison with the wild type and the mock-transduced cells, the proliferative response of the sense 6A8-transduced 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 6A8 alpha-mannosidase. Either enhancement or reduction of 6A8 alpha-mannosidase expression had no effect on proliferation induced by LMW-BCGF or IL-6.