腺病毒55型在A549细胞中的增殖动力学特征  

Proliferative Kinetics Characteristics of Adenovirus Type 55 in Human Lung Cell Line A549

作　　者：张绮玲[1,2] 李庆军[2] 王鑫[2] 曲新艳[2] 杨全[1] 杨静[2,3] 

机构地区：[1]广东药学院中药学院,广东广州510006 [2]军事医学科学院放射与辐射医学研究所,北京100850 [3]河南中医药大学药学院,郑州河南450008

出　　处：《生物技术通讯》2016年第1期48-51,共4页Letters in Biotechnology

基　　金：国家自然科学基金(31270197;81473184);国家重大新药创制课题(2013ZX09304102;2012ZX09301003-001-004);国家高技术研究发展计划(2012AA022501)

摘　　要：目的：利用噬斑法研究人呼吸道腺病毒55型（HAd V-B55）在A549细胞中的增殖动力学特征。方法：以正交法得出HAd V-B55的最佳噬斑条件;HAd V-B55以感染复数（MOI）为10感染A549细胞,噬斑法测定接种后3、6、9、12、15、18、21、24、36、48、60和72 h培养液上清中的病毒滴度,绘制log2（病毒滴度）-时间图,分析HAd V-B55在A549细胞中的增殖动力学特征。结果：终浓度含0.5%琼脂糖、2%胎牛血清和1640（2×）细胞培养液为HAd V-B55噬斑形成的最佳条件;MOI=10时,HAd V-B55感染A549细胞后12～15 h培养液中开始出现病毒,接种后15～36 h培养液中病毒量呈指数大量增加,接种后48～60 h病毒量出现小幅上升,接种后60 h细胞完全病变,病毒量增加达到平台期,最终病毒量增加约225 000倍。结论：A549细胞能有效地扩增HAd V-B55,其增殖周期为12～15 h;为研究HAd V-B55的感染致病机制,以及指导后期抗HAd V-B55的药物研发奠定了基础。Objective： To study the proliferative kinetics of human adenovirus type 55（HAdV-B55） in the hu- man lung cell line A549 using plaque assay. Methods： The orthogonal test was designed to obtain optimal medi- um for plaque assay. The A549 cells were infected by HAdV-B55 at a multiplicity of infection（MOI） of 10. Af- ter being infected for 1 h, the cells were overlaid with 1640 medium（2% FBS contained） and incubated at 37℃ under 50/0 CO2 incubator. The supernatants were separately harvested at 3, 6, 9, 12, 15, 18, 21, 24, 36, 48, 60 and 72 h post-infection and the proliferative kinetics of HAdV-B55 on A549 cells were determined by measuring titers of the supernatant samples using plaque assay. Results： The medium containing 0.5% agarose, 2% FBS and 1640（2x） was the optimal slipcover. When infected with HAdV-B55 at an MOI=IO, the virus began to appear in 12 to 15 h, the amount of virus increase exponentially in 15 to 36 h, the final viral in superuatant of 72 h was increased about 225 000 times. Conclusion： Proliferative kinetics analysis showed that HAdV-B55 could efficient- ly replicate and had a multiplication cycle of about 12～15 h in A549 cells. This work may contribute to the de- velopment of novel antiviral treatments for HAdV-B55 infection.

关 键 词：腺病毒55型 噬斑 增殖动力学 A549细胞 

分 类 号：R373[医药卫生—病原生物学] Q25[医药卫生—基础医学]