Yanagiya Akiko, Jia Qingmei, Ohka Seii, Horie Hitoshi, Nomoto Akio
Department of Microbiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
J Virol. 2005 Feb;79(3):1523-32. doi: 10.1128/JVI.79.3.1523-1532.2005.
The poliovirus (PV)-induced cytopathic effect (CPE) was blocked in neural cells but not in HeLa cells by the addition of monoclonal antibody (MAb) against PV or the human PV receptor (CD155) 2 h postinfection (hpi). Since each MAb has the ability to block viral infection, no CPE in PV-infected neural cells appeared to result from the blockade of multiple rounds of viral replication. Pulse-labeling experiments revealed that virus-specific protein synthesis proceeded 5 hpi with or without MAbs. However, in contrast to the results obtained without MAbs, virus-specific protein synthesis with MAbs was not detected 7 hpi. Shutoff of host translation was also not observed in the presence of MAbs. Western blot analysis showed that 2Apro, the viral protein which mediates the cleavage of eukaryotic translation initiation factor eIF4G, was still present 11 hpi. However, intact eIF4G appeared 11 hpi. An immunocytochemical study indicated that 2Apro was detected only in the nucleus 11 hpi. These results suggest that neural cells possess protective response mechanisms against PV infection as follows: (i) upon PV infection, neural cells produce a factor(s) to suppress PV internal ribosome entry site activity by 7 hpi, (ii) a factor which supports cap-dependent translation for eIF4G may exist in infected cells when no intact eIF4G is detected, and (iii) the remaining 2Apro is not effective in cleaving eIF4G because it is imported into the nucleus by 11 hpi.
感染后2小时(hpi)添加抗脊髓灰质炎病毒(PV)或人PV受体(CD155)的单克隆抗体(MAb),可阻断PV在神经细胞中诱导的细胞病变效应(CPE),但不能阻断其在HeLa细胞中的效应。由于每种MAb都有阻断病毒感染的能力,因此PV感染的神经细胞中未出现CPE似乎不是由于多轮病毒复制被阻断所致。脉冲标记实验显示,无论有无MAb,病毒特异性蛋白质合成在感染后5小时均能进行。然而,与未使用MAb的结果相反,在感染后7小时未检测到使用MAb时的病毒特异性蛋白质合成。在存在MAb的情况下也未观察到宿主翻译的关闭。蛋白质免疫印迹分析表明,介导真核翻译起始因子eIF4G裂解的病毒蛋白2Apro在感染后11小时仍然存在。然而,完整的eIF4G在感染后11小时出现。免疫细胞化学研究表明,在感染后11小时仅在细胞核中检测到2Apro。这些结果表明,神经细胞对PV感染具有如下保护反应机制:(i)在PV感染后,神经细胞会产生一种或多种因子,在感染后7小时抑制PV内部核糖体进入位点活性;(ii)当未检测到完整的eIF4G时,感染细胞中可能存在支持eIF4G帽依赖性翻译的因子;(iii)剩余的2Apro在感染后11小时因被导入细胞核而无法有效裂解eIF4G。