应用KMB17细胞培养柯萨奇病毒B族1、2、3型的研究  

Study on culture conditions of Coxsackievirus B type 1-3 on human embryonic lung diploid cell line KMB17

作　　者：邓自君[1] 靳玮华 李艳艳[1] 张伟 杨凤梅[1] 张名[1] 陈丽雄[1] 徐鸿界 李明学 刘权 唐洁 段素琴[1] 孙文亭 李咏洁 马绍辉[1] 赵远[1] 杨明 和占龙[1] 杨净思[1] DENG Zijun;JIN Weihua;LI Yanyan;ZHANG Wei;YANG Fengmei;ZHANG Ming;CHEN Lixiong;XU Hongjie;LI Mingxue;LIU Quan;TANG Jie;DUAN SUqing;SUN Wenting;LI Yongjie;MA Shaohui;ZHAO Yuan;YANG Ming;HE Zhanlong;YANG Jingsi(Institute of Medical Biology,Chinese Academy of Medical Sciences&Peking UnionMedical College,Kunming 650018,China)

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,云南昆明650018

出　　处：《中国病原生物学杂志》2024年第1期10-14,共5页Journal of Pathogen Biology

基　　金：国家重点研发计划项目(No.2021YFF0702804);中国医学科学院医学与健康科技创新工程项目(No.2021-I2M-1-024);中国医学科学院医学生物学研究所所级重点科研专项项目(No.2022IMBCAMS002);云南省重大科技专项计划项目(No.202002AA100009)。

摘　　要：目的 采用人胚肺二倍体细胞(human embryonic lung diploid cell)KMB17株对柯萨奇病毒B族(Coxsackievirus B,CVB)1、2、3型进行适应性培养,探索应用KMB17细胞培养CVB1-3型病毒的相关参数,为基于KMB17细胞研发CVB多价疫苗提供参考。方法 将CVB 1-3型分别设置8个不同的感染复数(mulitiplicity of infection, MOI)梯度(0.05、0.1、0.15、0.2、0.25、0.3、0.35、0.4),在KMB17细胞上进行培养观察,对病毒培养结果进行统计分析。结果 CVB 1-3型病毒均能KMB17细胞中复制,并使细胞产生明显的细胞病变(cytopathic effect, CPE)。其中CVB 1型在KMB17细胞上适宜的种毒MOI值为0.25~0.3,病毒收获液的平均滴度为7.25 lgCCID50/mL,适宜MOI范围内病毒培养时间的95%置信区间为96.1~97.6 h;CVB 2型在KMB17细胞上适宜的种毒MOI值为0.20~0.25,病毒收获液平均滴度为7.375 lgCCID50/mL,适宜MOI范围内病毒培养时间的95%置信区间为74.8~76.8 h。CVB 3型在KMB17细胞上适宜的种毒MOI值为0.30~0.35,病毒收获液的平均滴度为7.25 lgCCID50/mL,适宜MOI范围内病毒培养时间的95%置信区间为82.7~84.7 h。结论 KMB17为一株理想的用于CVB1、2、3型培养的基质细胞,可为基于KMB17细胞研发CVB多价疫苗提供参考。Objective To use human embryonic lung diploid cell KMB17 strain for adaptive culture of Coxsackievirus B(CVB)types 1,2,and 3,and to explore the relevant parameters of using KMB17 cells to culture CVB1-3 virus,providing reference for the development of CVB multivalent vaccines based on KMB17 cells.Methods CVB 1-3 types were cultured on KMB17 cells with 8 different mulitiplicity of infection(MOI)gradients(0.05,0.1,0.15,0.2,0.25,0.3,0.35,0.4),and the virus culture results were statistically analyzed.Results CVB 1-3 viruses can replicate in KMB17 cells and produce obvious cytopathic effect(CPE).The suitable MOI value for CVB type 1 virus on KMB17 cells is 0.25-0.3,the average titer of virus harvesting solution is 7.25 lgCCID_(50)/mL,and the 95%confidence interval for virus cultivation time within the appropriate MOI range is 96.1-97.6 hours;The suitable MOI value for CVB type 2 virus on KMB17 cells is 0.20-0.25,the average titer of virus harvest solution is 7.375 lgCCID_(50)/mL,and the 95%confidence interval for virus culture time within the suitable MOI range is 74.8-76.8 hours.The suitable MOI value for CVB type 3 virus on KMB17 cells is 0.30-0.35,the average titer of the virus harvest solution is 7.25 lgCCID_(50)/mL,and the 95%confidence interval for virus culture time within the appropriate MOI range is 82.7-84.7 hours.Conclusion KMB17 is an ideal stromal cell for culture of CVB1,2,and 3 types,which can provide reference for the development of CVB multivalent vaccines based on KMB17 cells.

关 键 词：KMB17细胞 柯萨奇病毒B 1、2、3型 病毒培养 

分 类 号：R373[医药卫生—病原生物学]