流感病毒疫苗株在MDCK细胞中增殖条件的优化  

Optimization of condition for propagation of influenza vaccine virus strain in MDCK cells

作　　者：李兴航 刘琛 张家友 闫璐瑶 韩天 刘博 李芳 宋颖丽 杨晓明 LI Xing-hang;LIU Chen;ZHANG Jia-you;YAN Lu-yao;HAN Tian;LIU Bo;LI Fang;SONG Ying-li;YANG Xiao-ming(Research Department of Viral Vaccine,Wuhan Institute of Biological Products Co.,Ltd.,Wuhan 430207,Hubei Province,China;不详)

机构地区：[1]武汉生物制品研究所有限责任公司,湖北武汉430207 [2]国家联合疫苗工程技术研究中心,湖北武汉430207 [3]中国生物技术股份有限公司,北京100029

出　　处：《中国生物制品学杂志》2021年第3期249-254,共6页Chinese Journal of Biologicals

基　　金：“重大新药创制”科技重大专项(2016ZX09106003-008)。

摘　　要：目的优化四价流感裂解疫苗中H1N1(IVR-180)、H3N2(IVR-186)、BV(NYMC BX-69A)、BY(B/Phuket/3073/2013)亚型疫苗株在MDCK细胞中的增殖条件。方法以血凝滴度和半数组织感染剂量(median tissue culture infective dose,TCID50)为指标,通过棋盘法对4种亚型流感病毒疫苗株在MDCK细胞上种毒的MOI(0.1、0.01、0.001、0.000 1)、TPCK胰酶终浓度(0.5、1、1.5、2、3、4μg/mL)及收毒时间(0、24、48、72、96 h)进行优化,并检测各条件下MDCK细胞的葡萄糖和乳酸代谢情况。结果 H1N1(IVR-180)、H3N2(IVR-186)、BV(NYMC BX-69A)、BY(B/Phuket/3073/2013)4种亚型流感病毒疫苗株的最适MOI分别为0.000 1、0.001、0.000 1和0.001,TPCK胰酶终浓度分别为1、2、1和2μg/mL,收毒时间分别为72、48、48和48 h。各条件下MDCK细胞的葡萄糖和乳酸代谢水平呈整体上升趋势。结论成功优化了4种亚型流感病毒疫苗株在MDCK细胞上的增殖条件,为以MDCK细胞为基质流感疫苗的研发奠定了基础。Objective To optimize the condition for propagation of influenza virus strains of four subtypes contained in quadrivalent split influenza vaccine,i.e. H1N1(IVR-180),H3N2(IVR-186),BV(NYMC BX-69A)and BY(B/Phuket/3073/2013),in MDCK cells. Methods The MOI(0. 1,0. 01,0. 001 and 0. 000 1),TPCK trypsin concentration(0. 5,1,1. 5,2,3 and 4 μg/mL)and virus harvest time(0,24,48,72 and 96 h)for inoculation and culture of four subtypes of influenza vaccine virus strains in MDCK cells were optimized by chessboard assay using the hemagglutination titer and median tissue culture infective dose(TCID50)as indicators. The glucose and lactic acid metabolisms of MDCK cells under various conditions were monitored. Results The optimal MOIs of influenza vaccine virus strains of subtypes H1N1(IVR-180),H3N2(IVR-186),BV(NYMC BX-69A)and BY(B/Phuket/3073/2013)were 0. 000 1,0. 001,0. 000 1 and 0. 001,while the final TPCK trypsin concentrations were 1,2,1 and 2 μ g/mL,and the harvest times were 72,48,48 and 48 h,respectively. Both the glucose and lactic acid metabolism levels under various conditions showed increasing trends in general. Conclusion The condition for propagation of four subtypes of influenza virus strains in MDCK cells was successfully optimized,which laid a foundation of development of influenza vaccine based on MDCK cells.

关 键 词：MDCK细胞 流感病毒 四价流感疫苗 增殖 

分 类 号：R373.13[医药卫生—病原生物学]