Epstein Barr Virus Induced Genomic Instability

描述（由申请人提供）：EB病毒（EBV）是一种 无处不在的人类疱疹病毒，尽管生活时间的关系， 与其人类宿主实现，可以与良性（传染性） 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 该基金的目的是了解EBV的分子机制， 导致疾病及其相互关系的模式，病毒的持久性， 记忆B淋巴细胞库。通过B细胞抗原的生理信号传导 受体（表面免疫球蛋白）对任何 感染的B细胞，导致细胞增殖和分化，或者， 相反，凋亡。因为我们发现重组酶的表达上调 激活基因RAG 1和RAG 2在EBV感染成熟B细胞后，我们现在 假设病毒通过以下途径使B细胞抗原受体多样化 诱导二级免疫球蛋白基因重排作为一种手段， 确保在受感染的细胞后代中有足够的存活信号。更新V（D）J 在骨髓或胚泡的选择性环境之外的重组 中心具有潜在的致病后果，包括自身免疫， 淋巴增生和染色体损伤。具体目的是测试我们的 假设是：1）确定免疫球蛋白二级重排是否 可变区基因的出现是爱泼斯坦-巴尔病毒诱导RAG的结果 病毒; 2）确定RAG 1和RAG 2是否在人外周血中表达 淋巴细胞在体内作为急性EBV感染的结果; 3）分析EBV DNA整合作为病毒引起的非法重组的标志 诱导RAG表达; 4）确定RAG 1和RAG 2 被潜伏蛋白EBNA 1上调。重组EBV的用途 表达绿色荧光蛋白允许快速选择感染的细胞 现在能够表达RAG;通过流式细胞术同时分析 改变的表面免疫球蛋白;通过PCR检测断裂的DNA末端或切除 环是V（D）J重组的副产品;以及随后的分析， RAG异常导致的染色体异常

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.