当前位置: X-MOL 学术Proc. Natl. Acad. Sci. U.S.A. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Human sapovirus propagation in human cell lines supplemented with bile acids [Microbiology]
Proceedings of the National Academy of Sciences of the United States of America ( IF 9.4 ) Pub Date : 2020-12-15 , DOI: 10.1073/pnas.2007310117
Hirotaka Takagi 1 , Tomoichiro Oka 2 , Takashi Shimoike 2 , Hiroyuki Saito 3 , Takayuki Kobayashi 4 , Tomoko Takahashi 5 , Chika Tatsumi 6 , Michiyo Kataoka 7 , Qiuhong Wang 8 , Linda J. Saif 8 , Mamoru Noda 9
Affiliation  

Human sapoviruses (HuSaVs) cause acute gastroenteritis similar to human noroviruses. Although HuSaVs were discovered four decades ago, no HuSaV has been grown in vitro, which has significantly impeded the understanding of viral biology and the development of antiviral strategies. In this study, we identified two susceptible human cell lines, that originated from testis and duodenum, that support HuSaV replication and found that replication requires bile acids. HuSaVs replicated more efficiently in the duodenum cell line, and viral RNA levels increased up to ∼6 log10-fold. We also detected double-stranded RNA, viral nonstructural and structural proteins in the cell cultures, and intact HuSaV particles. We confirmed the infectivity of progeny viruses released into the cell culture supernatants by passaging. These results indicate the successful growth of HuSaVs in vitro. Additionally, we determined the minimum infectious dose and tested the sensitivities of HuSaV GI.1 and GII.3 to heat and ultraviolet treatments. This system is inexpensive, scalable, and reproducible in different laboratories, and can be used to investigate mechanisms of HuSaV replication and to evaluate antivirals and/or disinfection methods for HuSaVs.



中文翻译:

人沙波病毒在补充了胆汁酸的人细胞系中繁殖[微生物学]

人沙波病毒(HuSaVs)引起的急性胃肠炎类似于人诺如病毒。尽管HuSaV在四十年前被发现,但尚未在体外培养出HuSaV,这极大地阻碍了对病毒生物学和抗病毒策略发展的了解。在这项研究中,我们鉴定了两种敏感的人类细胞系,它们来自睾丸和十二指肠,它们支持HuSaV复制,并发现复制需要胆汁酸。HuSaV在十二指肠细胞系中复制效率更高,病毒RNA水平增加至约6 log 10-折。我们还检测了细胞培养物中的双链RNA,病毒非结构和结构蛋白以及完整的HuSaV颗粒。我们证实了通过传代释放到细胞培养上清液中的子代病毒的感染性。这些结果表明HuSaVs在体外成功生长。此外,我们确定了最小感染剂量,并测试了HuSaV GI.1和GII.3对加热和紫外线处理的敏感性。该系统便宜,可扩展且可在不同的实验室中重现,可用于研究HuSaV复制的机制并评估HuSaV的抗病毒剂和/或消毒方法。

更新日期:2020-12-16
down
wechat
bug