Department of Molecular Virology, Baylor College of Medicine, Houston, Texas, USA.
mBio. 2012 Jul 3;3(4):e00159-12. doi: 10.1128/mBio.00159-12. Print 2012.
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.
现在已经可以将干细胞系定向分化为称为诱导人肠道类器官（iHIOs）的类似肠道的组织（J.R.斯彭斯，C.N.梅休，S.A.兰金，M.F.库哈尔，J.E.瓦兰斯，K.托勒，E.E.霍斯金斯，V.V.卡林尼琴科，S.I.威尔斯，A.M.佐恩，N.F.施罗耶尔和 J.M.威尔斯，《自然》470:105-109,2011）。我们测试了 iHIOs 作为一种新的模型来培养和研究粪便病毒。开发了用实验室株轮状病毒，猴 SA11 感染 iHIOs 的方案。原理验证分析表明，iHIOs 支持肠道病毒轮状病毒的复制，这是基于免疫荧光检测非结构病毒蛋白（非结构蛋白 4[NSP4]和 NSP2）、定量逆转录-PCR（qRT-PCR）检测病毒 RNA 水平增加和产生感染性后代病毒。iHIOs 还被证明可以直接从粪便样本中支持 12/13 种临床轮状病毒分离株的复制。一个意外的发现是，不仅在 iHIOs 的上皮细胞中，而且在间质细胞群中都检测到轮状病毒感染。这项工作表明，iHIOs 为研究轮状病毒和其他肠道病毒提供了一种很有前途的新模型。
胃肠道病毒感染是儿童和成人患病和死亡的主要原因。由于缺乏在实验室中培养许多胃肠道病毒的充分方法，因此完全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了阻碍。诱导的人肠道类器官（iHIOs）是生成类似肠道组织的一种很有前途的新模型。这是首次报道使用 iHIOs 培养任何微生物的研究，在这种情况下，是一种肠病毒。实验室和临床轮状病毒分离株都可以在 iHIOs 中复制的证据表明，该模型不仅对轮状病毒的研究有用，而且对其他可能的感染因子也有用。此外，在意想不到的细胞类型中检测到轮状病毒蛋白突出了该系统的前景，可以揭示以前在其他肠道细胞培养模型中未被认识或研究过的发病机制新问题。
