Infection of primary B cells by the human lymphomagenic pathogen HHV8

描述(申请人提供)：原发渗出性淋巴瘤(PEL)和多中心Castleman病出现在携带人类疱疹病毒8型(HHV8)的免疫受损患者中，人类疱疹病毒8型是卡波西肉瘤的相同致癌病原体。像EBV一样，HHV8通过潜伏感染B细胞建立终身感染，并表达能够调节B细胞信号和生存的病毒蛋白。然而，与EBV形成鲜明对比的是，KSHV感染的细胞在无症状的人中极其罕见，而且KSHV在体外有效地感染B细胞已被证明是困难的。这表明KSHV可能感染一小部分B细胞。这个项目的长期目标是了解初级B细胞感染的生物学和发病机制，验证目标B细胞可能代表一个独特的环境的假设，在免疫妥协的背景下，允许细胞转化。我们希望更好地了解这一过程的规则，我们最初的目标是：1.鉴定支持KSHV感染的B细胞亚群(S)。我们将利用高通量多模式成像技术识别KSHV感染的B细胞靶点。认识到感染可能改变B细胞表型，我们将通过在感染前分离B细胞亚群并比较每个亚群的感染效率来确认KSHV靶标的同一性。2.确定KSHV感染对B细胞存活和功能的影响，并确定导致这些变化的候选基因。我们将通过检测KSHV在体外的增殖能力、存活和转化能力来重点研究KSHV诱导的变化。在这些功能研究的同时，我们将使用专注于B细胞激活途径的微型微阵列，然后是定量RT-PCR和/或蛋白质印迹，以确定可能导致潜在表型变化的细胞信号通路的变化。与公共卫生的相关性：患者的癌症和免疫细胞疾病在世界各地导致显著的发病率和死亡率。不幸的是，控制这些疾病如何发生以及健康细胞如何转变为恶性细胞的规则仍然不清楚。通过在实验室中检查可能导致这些类型癌症的人类病毒，我们希望我们将深入了解这些规律，为生物医学科学家提供开发更好的治疗和预防方法的工具。

DESCRIPTION (provided by applicant): Primary effusion lymphoma (PEL) and multicentric Castleman's disease arise in immunocompromised patients harboring human herpesvirus 8 (HHV8), the same oncogenic pathogen underlying Kaposi's sarcoma. Like EBV, HHV8 establishes lifelong infection by latently infecting B cells and expresses viral proteins capable of modulating B cell signaling and survival. Yet, in stark contrast to EBV, KSHV-infected cells are extremely rare in asymptomatic individuals, and efficient in vitro infection of B cells with KSHV has proven difficult. This suggests that KSHV may infect a small subset of B cells. The long-term goals of this project are to understand the biology and pathogenesis underlying primary B cell infection, examining the hypothesis that the target B cells may represent a unique environment allowing, in the setting of immunocompromise, cellular transformation. We hope to better understand the rules governing this process, with our initial goals being to: 1. Identify the subset(s) of B cells that support KSHV infection. We will utilize high throughput multimodal imaging technology to identify B cell targets of KSHV infection. Appreciating that infection may alter the B cell phenotype, we will confirm the identity of KSHV targets by isolating B cell subsets prior to infection and comparing the efficiency of infection in each subset. 2. Determine the effects of KSHV infection on B cell survival and function and identify candidate genes responsible for these changes. We will focus initial studies on KSHV-induced changes by examining proliferative capacity, survival and transformation ability in vitro. In parallel to these functional studies, we will use mini-microarrays focused on B cell activation pathways, followed by quantitative RT-PCR and/or western blot, to identify changes in cell signaling pathways that may be responsible for potential phenotypic changes. PUBLIC HEALTH RELEVANCE: Cancers and diseases of the immune cells in patients lead to significant morbidity and mortality throughout the world. Unfortunately, the rules governing how these diseases arise and how a healthy cell transforms into a malignant one remain unclear. By examining in the laboratory human viruses that can cause these types of cancers, it is our hope that we will gain insights into these rules, providing biomedical scientist the tools with which to develop better therapies and preventions.