HOST IMMUNITY TO EBV INFECTION IN VITRO AND IN VIVO

EB病毒（EBV）是一种持久性疱疹病毒， 潜伏和重新激活，通常对感染者没有伤害 单独的. EBV感染和“永生化”或“生长转化”正常 体外休眠的人类B细胞驱使它们成为潜伏感染 增殖的淋巴母细胞 该病毒还发挥了核心作用， 几种人类疾病，包括几种B细胞瘤。 的目标 这个建议将调查正常B 细胞生物学和建立一个潜在的，持续的感染， EB病毒 具体来说，我们将讨论这方面的两个方面 关系 首先，我们将详细分析 模型B细胞活化基因（Blast-1）受到调节。 Blast-1是一种 粘附分子，仅在淋巴和骨髓中表达 细胞，这是由EBV感染上调。 我们将绘制最小的 启动子和负责组织特异性表达的序列。 我们将建立转录和mRNA表达增加的相对作用， EBV激活的稳定性。 如此界定的监管要素将 被映射。 然后将该机制与其他已知的机制进行比较。 活化剂包括IL-4、PMA、IL-1b和离子霉素。 二是 研究持续存在的病毒基因组形式之间的关系， 和受感染细胞的分化阶段。 我们特别 将研究细胞活化阶段在决定 无论病毒是作为游离体还是整合体持续存在。 病毒 将研究涉及附加体扩增的序列。 将对先前鉴定的整合位点进行克隆和测序， 研究整合机制。 最后，肿瘤细胞系将 通过荧光检测来检测整合的EBV基因组的存在， 原位杂交技术提供了一个明确的答案， 无论这些序列是否存在。

Epstein-Barr virus (EBV) is a persistent herpesvirus that is carried latently and reactivates, usually without harm to the infected individual. EBV infects and "immortalizes" or "growth transforms" normal resting human B cells in vitro driving them to become latently infected proliferating lymphoblasts. The virus also plays a central role in several human diseases, including several B cell neoplasias. The goal of this proposal will be to investigate the relationship between normal B cell biology and the establishment of a latent, persistent infection by Epstein-Barr virus. Specifically, we will address two aspects of this relationship. First, we will undertake a detailed analysis of how a model B cell activation gene (Blast-1) is regulated. Blast-1 is an adhesion molecule, restricted in its expression to lymphoid and myeloid cells, that is upregulated by EBV infection. We will map the minimal promoter and the sequences responsible for tissue specific expression. We will establish the relative roles of increased transcription and mRNA stabilization in activation by EBV. Regulatory elements so defined will be mapped. The mechanism will then be compared to that of other known activators including IL-4, PMA, IL-lb and ionomycin. Second, we will study the relationship between the form of the persistent viral genome and the differentiation stage of the infected cell. Specifically, we will investigate the role of cellular activation stage in determining whether the virus persists as an episome or integrated. The viral sequences involved in episomal amplification will be investigated. Previously identified integration sites will be cloned and sequenced to investigate the mechanism of integration. Lastly, tumor cell lines will be tested for the presence of integrated EBV genomes by the fluorescence in situ hybridization technique to provide a definitive answer as to whether such sequences are or are not present.