Epstein Barr Virus Induced Genomic Instability

描述(申请人提供)：爱泼斯坦-巴尔病毒是一种 无处不在的人类疱疹病毒，尽管他们一生都很融洽 与其人类宿主实现的，可与良性(传染性)相关 单核细胞增多症)和恶性(伯基特淋巴瘤、霍奇金淋巴瘤，原发性 中枢神经系统淋巴瘤)淋巴增生性疾病。整体而言 这项资助的目的是了解EBV感染的分子机制 致病原因及其与病毒持续模式的相互关系 记忆B淋巴细胞库。B细胞抗原的生理信号转导 受体(表面免疫球蛋白)对任何 感染B细胞，导致细胞增殖和分化或， 反之，则是细胞凋亡。因为我们发现重组酶的表达上调 激活基因RAG1和RAG2在EB病毒感染成熟B细胞中的作用 假设病毒通过多种途径使B细胞抗原受体多样化 诱导二次免疫球蛋白基因重排作为一种手段 确保在感染的细胞后代中有足够的生存信号。续订V(D)J 骨髓或生发细胞选择性环境外的重组 中心有潜在的致病后果，包括自身免疫， 淋巴增殖和染色体损伤。具体的目的是测试我们的 假设是：1)确定免疫球蛋白的二次重排 可变区基因是爱泼斯坦-巴尔诱导RAG的结果 病毒；2)确定RAG1和RAG2是否在人外周血中表达 急性EBV感染对体内淋巴细胞的影响；3)EBV分析 DNA整合作为病毒引发的非法重组的标志 诱导RAG表达；4)确定RAG1和RAG2的表达机制 被潜伏蛋白EBNA1上调。重组EBV的应用 表达绿色荧光蛋白可以快速选择感染细胞 现在能够表达RAG；通过流式细胞术同时分析 表面免疫球蛋白改变；用聚合酶链式反应检测DNA末端断裂或切除 作为V(D)J重组的副产品的圆；以及随后对 异常破布引起的染色体异常。

DESCRIPTION (provided by the applicant): The Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that, despite the life-time rapport typically achieved with its human host, can be associated with benign (infectious mononucleosis) and malignant (Burkitt's lymphoma, Hodgkin's lymphoma, primary central nervous system lymphoma) lymphoproliferative diseases. The overall objective of this grant is to understand molecular mechanisms by which EBV causes disease and their inter-relatedness to modes of viral persistence in the memory B lymphocyte reservoir. Physiologic signaling via the B cell antigen receptor (surface immunoglobulin) has major implications for the fate of any infected B cell, leading to cell proliferation and differentiation or, conversely, apoptosis. Because we showed up-regulation of recombinase activating genes RAG1 and RAG2 upon EBV infection of mature B cells, we now hypothesize that virus diversifies the B cell antigen receptor through induction of secondary immunoglobulin gene rearrangements as a means of assuring adequate survival signaling in infected cell progeny. Renewed V(D)J recombination outside the selective environment of bone marrow or germinal centers has potential pathogenic consequences that include auto-immunity, lymphoproliferation and chromosomal damage. The specific aims to test our hypothesis are: 1) to determine if secondary rearrangements of immunoglobulin variable region genes occur as a consequence of RAG induction by Epstein-Barr virus; 2) to determine if RAG1 and RAG2 are expressed in human peripheral blood lymphocytes in vivo as a consequence of acute EBV infection; 3) to analyze EBV DNA integration as a marker of illegitimate recombination prompted by viral induced RAG expression; 4) to determine the mechanism by which RAG1 and RAG2 are up regulated by latency protein EBNA1. The use of recombinant EBV expressing green fluorescent protein allows rapid selection of infected cells now capable of expressing RAG; concurrent analysis by flow cytometry for altered surface immunoglobulin; detection by PCR of broken DNA ends or excision circles that are byproducts of V(D)J recombination; and subsequent analysis for chromosomal abnormalities from aberrant RAG.