Stem cell-derived human intestinal organoids as an infection model for rotaviruses.
Stem cell-derived human intestinal organoids as an infection model for rotaviruses.
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作者：
Finkbeiner SR;Zeng XL;Utama B;Atmar RL;Shroyer NF;Estes MK
干细胞系定向分化成被称为诱导的人肠类器官（iHIO）的类胡萝卜素组织现在是可能的（J.R.斯宾塞角N. Mayhew，S. A.兰金，M. F. Kuhar，J. E. Vallance，K. Tolle，E. E.霍斯金斯，V. V. Kalinichenko，S. I.威尔斯，A. M. Zorn，N. F. Shroyer和J.M.威尔斯，自然470：105-109，2011）。我们测试了iHIOs作为培养和研究粪便病毒的新模型。开发了用轮状病毒的实验室毒株猿猴SA 11感染iHIO的方案。原理验证分析表明，iHIO支持胃肠道病毒轮状病毒的复制，基于通过免疫荧光检测非结构病毒蛋白（非结构蛋白4 [NSP 4]和NSP 2），通过定量逆转录-PCR（qRT-PCR）增加病毒RNA水平，以及产生感染性子代病毒。iHIO还显示支持直接从粪便样品复制12/13个临床轮状病毒分离株。一个意想不到的发现是不仅在上皮细胞中而且在iHIO的间充质细胞群中检测到轮状病毒感染。这项工作表明，iHIO为研究轮状病毒和其他胃肠道病毒提供了一个有前途的新模型。胃肠道病毒感染是儿童和成人疾病和死亡的主要原因。由于在实验室中培养许多胃肠道病毒的方法不足，充分了解病毒如何与人类肠道细胞相互作用以引起疾病的能力受到阻碍。诱导的人肠类器官（iHIO）是一种有前途的新模型，用于产生类精氨酸组织。这是第一份使用iHIO培养任何微生物的研究报告，在这种情况下，是肠道病毒。实验室和临床轮状病毒分离株都可以在iHIO中复制的证据表明，该模型不仅可用于轮状病毒的研究，还可用于其他感染因子的研究。此外，在意想不到的细胞类型中检测轮状病毒蛋白突出了该系统揭示先前在其他肠细胞培养模型中未被认识或研究的关于发病机制的新问题的前景。
Directed differentiation of stem cell lines into intestine-like tissue called induced human intestinal organoids (iHIOs) is now possible (J. R. Spence, C. N. Mayhew, S. A. Rankin, M. F. Kuhar, J. E. Vallance, K. Tolle, E. E. Hoskins, V. V. Kalinichenko, S. I. Wells, A. M. Zorn, N. F. Shroyer, and J. M. Wells, Nature 470:105-109, 2011). We tested iHIOs as a new model to cultivate and study fecal viruses. Protocols for infection of iHIOs with a laboratory strain of rotavirus, simian SA11, were developed. Proof-of-principle analyses showed that iHIOs support replication of a gastrointestinal virus, rotavirus, on the basis of detection of nonstructural viral proteins (nonstructural protein 4 [NSP4] and NSP2) by immunofluorescence, increased levels of viral RNA by quantitative reverse transcription-PCR (qRT-PCR), and production of infectious progeny virus. iHIOs were also shown to support replication of 12/13 clinical rotavirus isolates directly from stool samples. An unexpected finding was the detection of rotavirus infection not only in the epithelial cells but also in the mesenchymal cell population of the iHIOs. This work demonstrates that iHIOs offer a promising new model to study rotaviruses and other gastrointestinal viruses. Gastrointestinal viral infections are a major cause of illness and death in children and adults. The ability to fully understand how viruses interact with human intestinal cells in order to cause disease has been hampered by insufficient methods for growing many gastrointestinal viruses in the laboratory. Induced human intestinal organoids (iHIOs) are a promising new model for generating intestine-like tissue. This is the first report of a study using iHIOs to cultivate any microorganism, in this case, an enteric virus. The evidence that both laboratory and clinical rotavirus isolates can replicate in iHIOs suggests that this model would be useful not only for studies of rotaviruses but also potentially of other infectious agents. Furthermore, detection of rotavirus proteins in unexpected cell types highlights the promise of this system to reveal new questions about pathogenesis that have not been previously recognized or investigated in other intestinal cell culture models.