VIRAL TRANSCRIPTION IN EBV TRANSFORMED B CELLS

EBV 转化的 B 细胞中的病毒转录

• 批准号: 7349181
• 负责人: SAMUEL H SPECK
• 金额: $ 4.01万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-09 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7349181
• 关键词: cell line; gene expression; infection; memory; methylation; role

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long term goal is to understand how Epstein-Barr virus (EBV) gene expression is regulated during immortalizing latency. EBV establishes a life-long infection within the infected host, and is closely associated with the development of endemic Burkitt?s lymphoma, nasopharyngeal carcinoma, 30-50% of Hodgkin?s disease, and nearly half of the lymphomas that arise in immunosuppressed patients. Notably, EBV infection of peripheral resting B cells results in growth transformation resulting, ex vivo, in the generation of immortalized lymphoblastoid cell lines. Based on recent analyses of EBV infection in seropositive individuals, it seems likely that the ability of EBV to drive B cell proliferation, and subsequent differentiation, plays an important role in the dissemination of virus infected B cells and the establishment of a long-lived latency reservoir in memory B cells (in which there is very limited viral gene expression). Understanding how EBV regulates viral gene expression during immortalizing latency may ultimately provide strategies for interfering with this phase of the virus life cycle, which could interfere with the establishment of latency in the memory B cell compartment. We continue to focus analyses on identifying and characterizing cis-elements involved in regulated EBNA gene expression during the immortalizing latency program of EBV. Aim 1. Generation of a cottontop marmoset LCL immortalized with a packaging defective and replication null EBV to serve as an inducible reservoir for non-immortalizing EBV mutants. Aim 2. Generation and characterization of EBV harboring mutations in cis-elements thought to be involved in regulating EBNA gene transcription in EBV immortalized lymphoblastoid cell lines. Aim 3. Analysis of the requirements for Wp activity during initial stages of infection of primary B cells. Aim 4. Analysis of Wp methylation during the establishment and maintenance of immortalizing latency in B cells ? role of methylation in regulating Wp activity in lymphoblastoid cell lines.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。长期目标是了解 Epstein-Barr 病毒 (EBV) 基因表达在永生潜伏期是如何受到调节的。 EBV 在受感染宿主体内形成终生感染，并与地方性伯基特淋巴瘤、鼻咽癌、30-50% 霍奇金病以及近一半免疫抑制患者中出现的淋巴瘤的发展密切相关。值得注意的是，外周静息 B 细胞的 EBV 感染会导致生长转化，从而在体外产生永生化类淋巴母细胞系。根据最近对血清阳性个体中 EBV 感染的分析，EBV 驱动 B 细胞增殖和随后分化的能力似乎在病毒感染 B 细胞的传播和长寿命潜伏期的建立中发挥着重要作用。记忆 B 细胞中的储存库（其中病毒基因表达非常有限）。了解 EBV 在永生潜伏期如何调节病毒基因表达可能最终提供干扰病毒生命周期这一阶段的策略，这可能会干扰记忆 B 细胞区室中潜伏期的建立。我们继续重点分析鉴定和表征 EBV 永生潜伏期过程中参与调节 EBNA 基因表达的顺式元件。 目标 1. 产生具有包装缺陷和复制无效 EBV 的永生化棉顶狨猴 LCL，作为非永生化 EBV 突变体的诱导型储存库。目标 2. 带有顺式元件突变的 EBV 的生成和表征，这些突变被认为参与调节 EBV 永生化淋巴母细胞系中的 EBNA 基因转录。目的3.分析原代B细胞感染初期对Wp活性的需求。目标 4. B 细胞永生潜伏期建立和维持过程中 Wp 甲基化的分析？甲基化在调节淋巴母细胞系 Wp 活性中的作用。