Repression of Oral Virus by Interferon & Nuclear Bodies

干扰素抑制口腔病毒

• 批准号: 6344426
• 负责人: Jerry L Taylor
• 金额: $ 23.63万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6344426
• 关键词: Vero cells; cell line; disease /disorder prevention /control; gene expression; gene induction /repression; genetic transcription; herpes simplex virus 1; immunogenetics; immunoprecipitation; immunoregulation; interferons; mouth; protein protein interaction; protein structure function; secondary infection; tissue /cell culture; transcription factor; virus DNA; virus infection mechanism; virus replication; western blottings; yeast two hybrid system

DESCRIPTION: (Provided by Applicant) Patients with AIDS resulting from HIV have essentially no cell-mediated immunity. Therefore, their innate immune responses become critical for protection from secondary infections. Members of the herpesvirus family including HSV, CMV, EBV, KSHV, and VZV are major contributors to secondary infections in the oral cavity of AIDS patients. Interferons are the primary antiviral cytokine produced by virus infected cells. IFNs play a major role in the innate antiviral immune response, repressing viral gene expression very early in infection, presumably at the nuclear sites of viral genome deposition, adjacent to the ND10 nuclear body. IFNs induce expression of two families of nuclear proteins, namely PML and Sp100, which associate with the ND10s and appear to participate in repression of viral gene transcription. To study the mechanisms of this inhibition, members of the PML and SP100 families of IFN-inducible nuclear body proteins will be characterized in transcription repression assays for inhibition of basal and viral transactivator-induced gene expression. In addition, two newly described proteins, namely MZF1B and RAZ1, which form nuclear complexes termed REMs, which repress activity will be examined. Using both transient and stably transfected cells, the specificity of the repressors, used alone or in combination will be examined using chemiluminescence assays for reporter gene expression. Nuclear localization of the repressors and co-localization with ND10 structures will be examined in cells treated with IFN, in cells infected with HSV, and in the absence of treatment or infection. To understand the mechanisms of action of repressors, proteins with which they associate will be identified by co-immunoprecipitation assays and yeast two-hybrid analysis. The association of REM proteins with a major transcriptional activator of HSV will be characterized. The IFN-responsiveness of REM proteins will be measured. The localization of viral DNA with the novel nuclear structures will be determined. Understanding specificity of the repression of viral gene transcription by IFN-induced and other nuclear proteins and the mechanisms by which repression occurs should provide insight into how the cell combats viral infection. This understanding should allow the development of strategies for treatment of secondary viral infections in the oral cavity of HIV-infected individuals that enhance or mimic these repressive activities.

描述：（由申请人提供）艾滋病毒引起的艾滋病患者 本质上没有细胞介导的免疫力。因此，他们先天免疫反应 对保护免受继发感染至关重要。成员 HSV，CMV，EBV，KSHV和VZV在内的疱疹病毒家族是主要的 艾滋病患者口腔中继发感染的贡献者。 干扰素是由病毒感染产生的主要抗病毒细胞因子 细胞。 IFNS在先天抗病毒免疫反应中起主要作用， 在感染中很早就抑制病毒基因表达，大概是在 病毒基因组沉积的核部位，与ND10核体相邻。 IFN诱导两个核蛋白质家族的表达，即PML和 SP100，与ND10相关联，似乎参与了压抑 病毒基因转录。为了研究这种抑制的机制， IFN诱导核体蛋白的PML和SP100家族的成员 将以转录抑制测定的抑制作用来表征 基底和病毒式反式激活诱导的基因表达。此外，两个新近 描述了形成称为核复合物的蛋白质，即MZF1B和RAZ1 REMS，将检查哪些压制活动。同时使用瞬态和稳定 转染的单元，阻遏物的特异性，单独使用或在 将使用报告基因的化学发光测定法检查组合 表达。阻遏物的核定位和与共同定位 ND10结构将在用IFN处理的细胞中检查，在感染的细胞中 使用HSV，在没有治疗或感染的情况下。理解 阻遏物的作用机制，与之关联的蛋白质的作用机制 通过共免疫沉淀测定法和酵母两杂化分析确定。这 REM蛋白与HSV的主要转录激活剂的关联将 被描述。将测量REM蛋白的IFN反应性。这 将确定病毒DNA与新型核结构的定位。 理解抑制病毒基因转录的特异性 IFN诱导的和其他核蛋白以及抑制的机制 发生应提供有关细胞如何打击病毒感染的洞察力。这 理解应允许制定治疗策略 艾滋病毒感染者口腔中的继发病毒感染 增强或模仿这些压制活动。