Luplertlop Natthanej, Suwanmanee San, Muangkaew Watcharamat, Ampawong Sumate, Kitisin Thitinan, Poovorawan Yong
Department of Microbiology and Immunology, Mahidol University, Bangkok.
Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Bangkok.
J Vector Borne Dis. 2017 Jul-Sep;54(3):207-214. doi: 10.4103/0972-9062.217611.
BACKGROUND & OBJECTIVES: An increase in Zika virus (ZIKV) epidemic during the last decade has become a major global concern as the virus affects both newborns and adult humans. Earlier studies have shown the impact of ZIKV infection in developing human foetus. However, effective in vitro model of target cells for studying the ZIKV infection in adult human neurons is not available. This study aimed to establish the use of human neuroblastoma cell line (SH-SY5Y) for studying an infection of ZIKV in vitro.
ZIKV growth kinetics, viral toxicity, and SH-SY5Y cell vialibity were determined after ZIKV infection in SH-SY5Y cells in vitro. ZIKV-infected SH-SY5Y cells were morphologically analysed and compared with nonhuman primate Vero cells. Furthermore, the susceptibility of SH-SY5Y cells to ZIKV infection was also determined.
The results showed that ZIKV efficiently infects SH-SY5Y cell lines in vitro. Gradual changes of several cellular homeostasis parameters including cell viability, cytotoxicity, and cell morphology were observed in ZIKVinfected SH-SY5Y cells when compared to mock-treated or non-human primate cells. Interestingly, ZIKV particles were detected in the nucleoplasmic compartment of the infected SH-SY5Y cells.
INTERPRETATION & CONCLUSION: The results suggest that ZIKV particle can be detected in the nucleoplasmic compartment of the infected SH-SY5Y cells beside the known viral replicating cytoplasmic area. Hence, SH-SY5Y cells can be used as an in vitro adult human neuronal cell-based model, for further elucidating the ZIKV biology, and highlight other possible significance of Zika virus distribution through nuclear localization, which may correlate to the neuropathological defects in ZIKV-infected adult humans.
过去十年间寨卡病毒(ZIKV)疫情的加剧已成为全球主要关注点,因为该病毒会影响新生儿和成年人。早期研究已表明ZIKV感染对发育中的人类胎儿的影响。然而,目前尚无用于研究ZIKV在成人神经元中感染的有效体外靶细胞模型。本研究旨在建立利用人神经母细胞瘤细胞系(SH-SY5Y)进行ZIKV体外感染研究的方法。
在体外将ZIKV感染SH-SY5Y细胞后,测定ZIKV的生长动力学、病毒毒性以及SH-SY5Y细胞活力。对ZIKV感染的SH-SY5Y细胞进行形态学分析,并与非人类灵长类动物的Vero细胞进行比较。此外,还测定了SH-SY5Y细胞对ZIKV感染的易感性。
结果表明,ZIKV能在体外有效感染SH-SY5Y细胞系。与模拟处理的细胞或非人类灵长类动物细胞相比,在ZIKV感染的SH-SY5Y细胞中观察到包括细胞活力、细胞毒性和细胞形态在内的几个细胞稳态参数的逐渐变化。有趣的是,在感染的SH-SY5Y细胞的核质区室中检测到了ZIKV颗粒。
结果表明,除了已知的病毒复制细胞质区域外,在感染的SH-SY5Y细胞的核质区室中也能检测到ZIKV颗粒。因此,SH-SY5Y细胞可作为基于成人神经元细胞的体外模型,用于进一步阐明ZIKV生物学特性,并突出寨卡病毒通过核定位分布的其他可能意义,这可能与ZIKV感染成人的神经病理学缺陷相关。