印度尼西亚基孔肯雅病毒分离株在多种体外人细胞系中的生长特性

O. N. Turnip, Rahma F. Hayati, Rizka Alawiyah, B. Yohan, Dionisius Denis, A. Bowolaksono, A. Soebandrio, R. T. Sasmono
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引用次数: 1

摘要

基孔肯雅热(CHIK)是一种由基孔肯雅病毒(CHIKV)感染引起的发热性疾病,是影响世界热带和亚热带地区人群的蚊媒病毒性疾病之一。该病的发病机制尚未完全阐明,对CHIKV的研究采用了多种方法,包括利用细胞系在体外研究CHIKV的生物学特性。为了评估人类细胞系模型对CHIK研究的适用性,我们将包括A549、Huh7和HepG2在内的多种人类细胞系感染CHIKV并测定其感染易感性。采用MTT法和空斑法分别测定细胞活力和病毒生长动力学。采用荧光活化细胞分选(FACS)和免疫荧光法测定系统中感染细胞的比例及其形态可视化。A549和Huh7人类细胞系在感染后均表现出稳定的高细胞活力,而与Vero-CCL81(一种常规用于其他虫媒病毒研究的猴子细胞系)相比,这些细胞中的CHIKV生长动力学显著降低。有趣的是,我们在HepG2人细胞系中观察到明显不同的结果,其中细胞活力和CHIKV生长动力学明显更高。流式细胞术和免疫荧光检测证实,HepG2细胞的CHIKV感染率高于A549人细胞株。我们得出结论,人肝细胞HepG2细胞系对CHIKV亚洲基因型易感,并提出了作为常用的A549和Vero细胞体外CHIKV研究的替代细胞。
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Growth Characteristics of Chikungunya Virus Isolate from Indonesia in Various Human Cell Lines in vitro
Chikungunya (CHIK) fever, a febrile illness caused by Chikungunya virus (CHIKV) infection, is one of mosquito-borne viral diseases affecting people living in the tropical and subtropical regions in the world. The pathogenesis of the disease is yet to be completely unraveled, and research on CHIK has been conducted by employing various methods, including using cell lines to investigate the biological characteristics of CHIKV in vitro. To assess the suitability of human cell line model for CHIK study, various human cell lines including A549, Huh7, and HepG2 were infected with CHIKV and assayed for their susceptibility to infection. The MTT and plaque assay methods were performed to measure cell viability and virus growth kinetics, respectively. Fluorescence-activated Cell Sorting (FACS) and immunofluorescence assay were performed to measure the proportion of infected cells in the system and their morphological visualization. Both A549 and Huh7 human cell lines showed stable high cell viability upon infection while CHIKV growth kinetics were significantly lower in these cells compared to Vero-CCL81, a monkey cell line that is routinely used in other arboviruses research. Interestingly, we observed significantly different results in HepG2 human cell line, in which cell viability and CHIKV growth kinetics were significantly higher. FACS and immunofluorescence assay confirm the higher infection rate of CHIKV in HepG2 than A549 human cell line. We concluded herethat human hepatocytes HepG2 cell line was susceptible to Asian Genotype of CHIKV and proposed as an alternative cell for the in vitro CHIKV studies to the commonly used A549 and Vero cells.
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