Zhirnov O P, Vorob'eva I V, Safonova O A, Malyshev N A, Schwalm F, Klenk H -D
Vopr Virusol. 2013 Jul-Aug;58(4):20-8.
The propagation of the pandemic influenza virus H1N1 in cultures of bronchial (Calu-3) and intestinal (Caco-2) differentiated epithelial cells of human origin was studied. The canine epithelial cell lines, MDCK-H and MDCK-2, were comparatively tested. The two human cell lines were found to be highly sensitive to the influenza pandemic strains A/Hamburg/05/09 and A/Moscow/501/2011 and maintained their replication without addition of trypsin to culture medium. Virus strains of seasonal influenza H1N1, such as A/Moscow/450/2003, A/Memphis/14/96, and laboratory strain A/PR/8/34, multiplied in these human cells in similar manner. The intracellular cleavage HA0-->HA1+HA2 by the host virus-activating protease (IAP) occurred in both human cell lines under infection with each influenza virus H1N1 including pandemic ones. Comparatively, this cleavage of all influenza H1N1 virus strains appeared to be either undetectable or low-detectible in MDCK-H and MDCK-2, respectively, thereby implying low levels of active IAP in these cells. Multiplication of pandemic and seasonal influenza H1N1 viruses in Calu-3 and Caco-2 cells caused cytopathic effect, which was accompanied with low autophagy and apoptosis events. These data allow recommending human cell lines, Calu-3 and Caco-2, for optimized isolation and passaging of clinical strains of Influenza pandemic viruses H1N1.
研究了甲型H1N1流感大流行病毒在人源支气管（Calu-3）和肠道（Caco-2）分化上皮细胞培养物中的增殖情况。对犬上皮细胞系MDCK-H和MDCK-2进行了对比测试。发现这两种人细胞系对甲型流感大流行毒株A/汉堡/05/09和A/莫斯科/501/2011高度敏感，在培养基中不添加胰蛋白酶的情况下仍能维持其复制。季节性甲型H1N1流感病毒株，如A/莫斯科/450/2003、A/孟菲斯/14/96和实验室毒株A/PR/8/34，在这些人细胞中的增殖方式相似。在感染包括大流行毒株在内的每种甲型H1N1流感病毒时，宿主病毒激活蛋白酶（IAP）在这两种人细胞系中均会发生细胞内HA0裂解为HA1 + HA2的过程。相比之下，在MDCK-H和MDCK-2细胞中，所有甲型H1N1流感病毒株的这种裂解分别似乎无法检测到或检测水平很低，这意味着这些细胞中活性IAP的水平较低。甲型H1N1流感大流行病毒和季节性流感病毒在Calu-3和Caco-2细胞中的增殖会引起细胞病变效应，同时伴有低水平的自噬和凋亡事件。这些数据表明，人细胞系Calu-3和Caco-2可用于优化甲型H1N1流感大流行病毒临床毒株的分离和传代。
