Bale J F, O'Neil M, Wentzien J, Schelper R
J Neuropathol Exp Neurol. 1987 Jan;46(1):72-83. doi: 10.1097/00005072-198701000-00007.
We used cultures of reaggregate embryonic mouse brain cells to study murine cytomegalovirus (MCMV) infection of neural tissues. After 21 to 28 days in culture, aggregates were infected with MCMV and studied sequentially for 14 days using virus assay, electron microscopy and indirect immunofluorescence. Infectious virus could be recovered from aggregate cultures beginning three days after infection, and peak virus titers were observed on day 7 in aggregate tissues and on day 14 in culture fluids. By transmission electron microscopy, intranuclear viral nucleocapsids were identified in neural cells at the periphery of the aggregates on day 3. Infection then spread centripetally into aggregate tissues so that by day 14 the majority of neural cells contained intranuclear inclusions, numerous nucleocapsids, and mature virus particles. Virion production in neural cells was the result of a sequence of events that included budding of nucleocapsids from the nucleus and envelopment of cytoplasmic virus particles by membranes of the Golgi apparatus. These studies indicate that MCMV infection of murine brain aggregate cultures is a potentially useful in vitro system for the study of CMV infections of neural tissue.
我们利用重新聚集的胚胎小鼠脑细胞培养物来研究神经组织的鼠巨细胞病毒(MCMV)感染。培养21至28天后,将聚集物用MCMV感染,并使用病毒测定、电子显微镜和间接免疫荧光法连续研究14天。感染后三天开始,可从聚集物培养物中回收感染性病毒,在聚集组织中第7天观察到病毒滴度峰值,在培养液中第14天观察到病毒滴度峰值。通过透射电子显微镜,在第3天在聚集物周边的神经细胞中鉴定出核内病毒核衣壳。然后感染向心扩散到聚集组织中,以至于到第14天,大多数神经细胞含有核内包涵体、大量核衣壳和成熟病毒颗粒。神经细胞中的病毒粒子产生是一系列事件的结果,这些事件包括核衣壳从细胞核出芽以及细胞质病毒颗粒被高尔基体膜包裹。这些研究表明,鼠脑聚集物培养物的MCMV感染是用于研究神经组织CMV感染的潜在有用的体外系统。