昏睡性脑炎病因的肠道病毒证据。
Evidence for an enterovirus as the cause of encephalitis lethargica.
机构信息
Faculty of Life Sciences, London Metropolitan University, London, UK.
出版信息
BMC Infect Dis. 2012 Jun 20;12:136. doi: 10.1186/1471-2334-12-136.
BACKGROUND
The epidemic of encephalitis lethargica (EL), called classical EL, was rampant throughout the world during 1917-1926, affecting half a million persons. The acute phase was lethal for many victims. Post-encephalitic parkinsonism (PEP) affected patients for decades. Our purpose was to investigate the cause of classical EL by studying the few available brain specimens. Cases of PEP and modern EL were also studied. Transmission electron microscopy (TEM) and immunohistochemistry were employed to examine brain from four classical EL cases, two modern EL cases and one PEP case.
METHODS
Standard methods for TEM, immunohistochemistry and RTPCR were applied.
RESULTS
27 nm virus-like particles (VLP) were observed in the cytoplasm and nuclei of midbrain neurons in all classical EL cases studied. Large (50 nm) VLP and 27 nm intranuclear VLP were observed in the modern EL cases and the PEP case. Influenza virus particles were not found. VLP were not observed in the control cases. TEM of cell cultures inoculated with coxsackievirus B4 and poliovirus revealed both small and large intranuclear virus particles and small cytoplasmic particles, similar to the VLP in EL neurons. In the EL brains, nascent VLP were embedded in putative virus factories and on endoplasmic reticulum (ER). The VLP in the cases of classical EL survived, whereas ribosomes underwent autolysis due to the lack of refrigeration and slow formaldehyde fixation of whole brain. The VLP were larger than ribosomes from well preserved brain. Immunohistochemistry of classical EL cases using anti-poliovirus and anti-coxsackievirus B polyclonal antibodies showed significant staining of cytoplasm and nuclei of neurons as well as microglia and neuropil. Purkinje cells were strongly stained.A 97-bp RNA fragment of a unique virus was isolated from brain tissue from acute EL case #91558. Sequence analysis revealed up to 95% identity to multiple human Enteroviruses. Additional cases had Enterovirus positive reactions by real time PCR.
CONCLUSIONS
The data presented here support the hypothesis that the VLP observed in EL tissue is an Enterovirus.
背景
昏睡性脑炎（EL）的流行，也被称为经典 EL，于 1917 年至 1926 年期间在全球范围内肆虐，影响了 50 多万人。急性阶段对许多受害者来说是致命的。脑炎后帕金森病（PEP）影响了患者数十年。我们的目的是通过研究少数可用的脑组织标本来研究经典 EL 的病因。还研究了现代 EL 和 PEP 的病例。采用透射电镜（TEM）和免疫组化技术研究了来自四个经典 EL 病例、两个现代 EL 病例和一个 PEP 病例的脑标本。
方法
应用 TEM、免疫组化和 RT-PCR 标准方法。
结果
在所有研究的经典 EL 病例的中脑神经元细胞质和核内观察到 27nm 病毒样颗粒（VLP）。在现代 EL 病例和 PEP 病例中观察到大（50nm）VLP 和 27nm 核内 VLP。未发现流感病毒颗粒。对照病例中未观察到 VLP。接种柯萨奇病毒 B4 和脊髓灰质炎病毒的细胞培养物的 TEM 显示出小和大的核内病毒颗粒和小的细胞质颗粒，类似于 EL 神经元中的 VLP。在 EL 脑内，新生的 VLP 被嵌入假定的病毒工厂和内质网（ER）中。由于缺乏冷藏和整个大脑的甲醛固定缓慢，经典 EL 病例中的 VLP 存活下来，而核糖体由于缺乏冷藏和整个大脑的甲醛固定缓慢而自溶。VLP 比来自保存完好的大脑的核糖体大。使用抗脊髓灰质炎病毒和抗柯萨奇病毒 B 多克隆抗体对经典 EL 病例进行免疫组化染色显示，神经元的细胞质和细胞核以及小胶质细胞和神经胶均有明显染色，浦肯野细胞染色强烈。从急性 EL 病例#91558 的脑组织中分离出一段 97bp 的独特病毒 RNA 片段。序列分析显示，该片段与多种人类肠道病毒的同源性高达 95%。其他病例通过实时 PCR 检测到肠道病毒阳性反应。
结论
本文提供的数据支持这样一种假设，即在 EL 组织中观察到的 VLP 是一种肠道病毒。
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