REGULATION OF VIRAL INFECTIONS BY CELLULAR GENES

细胞基因对病毒感染的调控

• 批准号: 2273319
• 负责人: J. Marie Hardwick
• 金额: $ 22.15万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-02-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2273319
• 关键词: SCID mouse; Sindbis virus; antioxidants; apoptosis; genetic strain; glycoproteins; host organism interaction; infectious encephalitis; laboratory mouse; neurotropic virus; oncogenes; oxidation reduction reaction; oxidative stress; protein folding; protein structure function; spinal ganglion; tissue /cell culture; transfection; virulence; virus RNA; virus genetics; virus infection mechanism; virus protein; virus replication

Alphaviruses cause encephalitis in man and animals and are transmitted by mosquitos. The alphavirus Sindbis infects and kills most cultured cell types, not by lethal parasitism, but by inducing programmed cell death, or apoptosis. However, in mosquito cells and in neurons, both in vivo and in vitro, Sindbis virus can establish a persistent infection in which virus continues to be produced but the cells survive. These results indicate that a cellular factor can modulate the outcome of infection. Neurons express the human oncogene bcl-2 which has been shown to block apoptosis in a variety of cell types and species. We generated a cell line expressing bcl-2 and found that it supported Sindbis virus persistence while controls lacking bcl-2 died. However, neurovirulent strains of Sindbis virus were able to induce apoptosis despite the expression of bcl-2. This ability to kill bcl2-containing cells has been mapped to a single amino acid change in the E2 glycoprotein. To study Sindbis virus neurovirulence, we will examine the molecular mechanisms by which viral E2 and cellular bcl-2 modulate the outcome of Sindbis virus infection. To determine why avirulent strains fail to kill the cells expressing bcl-2, we will identify the viral infection step (binding, entry, replication, maturation) which is affected. A battery of antibodies will be used study the maturation of E2 in bcl-2 positive and negative cells. In addition we will generate cell lines expressing bcl-2 mutants and assay the function of these mutants in Sindbis virus infections. Some effects of bcl-2 can be mimicked by treatment of cells with antioxidants. Therefore, we will study the effects of these agents on viral replication and maturation, and on the structure/function of the E2 glycoprotein. The mechanisms of Sindbis virus neurovirulence are likely to apply to other viruses, and an increased understanding of the bcl-2 oncogene has ramifications for normal cellular processes as well as neurodegenerative disease.

α病毒引起人和动物的脑炎，并传播 由蚊子。 alphavirus sindbis感染并杀死大多数培养的细胞 类型不是致命的寄生虫，而是通过诱导编程的细胞死亡， 或凋亡。但是，在蚊子细胞和神经元中，体内和 在体外，辛德比斯病毒可以建立一种持续感染，其中 病毒继续产生，但细胞存活。这些结果 表明细胞因子可以调节感染的结果。 神经元表达人类癌基型BCL-2，已证明可以阻止 各种细胞类型和物种的凋亡。我们生成了一个单元 表达Bcl-2的线并发现它支持信德氏病毒 持久性虽然缺乏BCL-2的控件死亡。然而，神经脊髓 尽管有病毒菌株，但尽管有 Bcl-2的表达。这种杀死含Bcl2细胞的能力已经 映射到E2糖蛋白中的单个氨基酸变化。 为了研究sindbis病毒神经动力学，我们将检查分子 病毒E2和细胞Bcl-2调节结果的机制 信德氏病毒感染。确定为什么无毒菌株无法杀死 表达Bcl-2的细胞，我们将确定病毒感染步骤 （绑定，进入，复制，成熟），受影响。电池 将使用抗体研究Bcl-2阳性中E2的成熟 和负细胞。另外，我们将生成表达细胞系 Bcl-2突变体并测定这些突变体在信德氏病毒中的功能 感染。 Bcl-2的某些效果可以通过细胞处理来模仿 抗氧化剂。因此，我们将研究这些试剂的影响 关于病毒复制和成熟，以及有关的结构/功能 E2糖蛋白。 信德氏病毒神经电动机的机制可能适用于 其他病毒以及对Bcl-2癌基因的了解已有 正常细胞过程以及神经退行性的后果 疾病。