VIRAL TRANSCRIPTION IN EBV TRANSFORMED B CELLS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。长期目标是了解eb病毒（EBV）基因表达在永生化潜伏期是如何被调控的。EBV在被感染的宿主体内建立终身感染，并与地方性伯基特病的发展密切相关。s淋巴瘤，鼻咽癌，30-50%霍奇金？在免疫抑制患者中出现的淋巴瘤中，有近一半是这种疾病。值得注意的是，EBV感染外周静止B细胞导致生长转化，从而在体外产生永生化淋巴母细胞样细胞系。根据最近对EBV感染血清阳性个体的分析，EBV驱动B细胞增殖和随后分化的能力似乎在病毒感染B细胞的传播和在记忆B细胞中建立长寿命潜伏库（其中病毒基因表达非常有限）中发挥重要作用。了解EBV如何在永生化潜伏期调节病毒基因表达，可能最终提供干扰病毒生命周期这一阶段的策略，从而干扰记忆B细胞区室中潜伏期的建立。在EBV永生化潜伏期程序中，我们将继续专注于识别和表征参与EBNA基因表达调控的顺式元件。目的1。用包装缺陷和复制无效的EBV永生化棉绒猴LCL的一代，作为非永生化EBV突变体的诱导库。目标2。在EBV永生化淋巴母细胞系中，被认为参与调节EBNA基因转录的顺式元件突变的EBV的产生和特性。目标3。原代B细胞感染初期对Wp活性的需求分析。目标4。B细胞永生化潜伏期建立和维持过程中Wp甲基化的分析？甲基化在淋巴母细胞样细胞系中调节Wp活性的作用。

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.