Ng Ching G, Griffin Diane E
W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205, USA.
J Virol. 2006 Nov;80(22):10989-99. doi: 10.1128/JVI.01154-06. Epub 2006 Aug 30.
Sindbis virus (SV), an enveloped virus with a single-stranded, plus-sense RNA genome, is the prototype alphavirus in the Togaviridae family. In mice, SV infects neurons and can cause apoptosis of immature neurons. Sphingomyelin (SM) is the most prevalent cellular sphingolipid, is particularly abundant in the nervous systems of mammals, and is required for alphavirus fusion and entry. The level of SM is tightly regulated by sphingomyelinases. A defect in acid sphingomyelinase (ASMase) results in SM storage and subsequent intracellular accumulation of SM. To better understand the role of the SM pathway in SV pathogenesis, we have characterized SV infection of transgenic mice deficient in the ASMase gene. ASMase knockout (ASM-KO) mice were more susceptible to SV infection than wild-type (WT) or heterozygous (Het) animals. Titers of SV were higher in the brains of ASM-KO mice than in the brains of WT mice. More SV RNA was detected by in situ hybridization, more SV protein was detected by immunohistochemistry, and more terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling-positive cells were present in the cortex and hippocampus of ASM-KO mice than in those of WT or Het mice. Interleukin-6 (IL-6), but not IL-1beta or tumor necrosis factor alpha, was elevated in infected ASM-KO mice compared to levels in WT or Het mice, but studies with IL-6-KO mice and recombinant SV expressing IL-6 showed no role for IL-6 in fatal disease. Together these data indicate that the increase in susceptibility of ASM-KO mice to SV infection was the result of more-rapid replication and spread of SV in the nervous system and increased neuronal death.
辛德毕斯病毒(SV)是一种具有单链正义RNA基因组的包膜病毒,是披膜病毒科中甲病毒属的原型病毒。在小鼠中,SV感染神经元并可导致未成熟神经元凋亡。鞘磷脂(SM)是最普遍的细胞鞘脂,在哺乳动物神经系统中含量尤其丰富,是甲病毒融合和进入所必需的。SM的水平受到鞘磷脂酶的严格调控。酸性鞘磷脂酶(ASMase)缺陷会导致SM储存以及随后SM在细胞内积累。为了更好地理解SM途径在SV发病机制中的作用,我们对ASMase基因缺陷的转基因小鼠的SV感染进行了特征分析。与野生型(WT)或杂合子(Het)动物相比,ASMase基因敲除(ASM-KO)小鼠对SV感染更易感。ASM-KO小鼠脑中的SV滴度高于WT小鼠脑中的滴度。通过原位杂交检测到ASM-KO小鼠大脑中更多的SV RNA,通过免疫组织化学检测到更多的SV蛋白,并且与WT或Het小鼠相比,ASM-KO小鼠的皮质和海马中存在更多的末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记阳性细胞。与WT或Het小鼠相比,感染的ASM-KO小鼠中白细胞介素-6(IL-6)升高,但IL-1β或肿瘤坏死因子α未升高,但对IL-6基因敲除小鼠和表达IL-6的重组SV的研究表明IL-6在致命疾病中不起作用。这些数据共同表明,ASM-KO小鼠对SV感染易感性增加是SV在神经系统中更快复制和传播以及神经元死亡增加的结果。