AIDS LYMPHOMA HARBORING TWO GAMMA HERPESVIRUSES

描述（改编自研究者摘要）：本研究 集中在KSHV，它已被发现在KS和一些活检 非霍奇金淋巴瘤BCBL系，其潜伏感染有 KSHV和EBV，是唯一已知的KSHV生长的细胞培养来源 在细胞培养中，以及唯一已知的人类细胞， 感染了两种人类疱疹病毒初步研究 证明BCBL系可被诱导表达KSHV裂解性 循环肽和扩增KSHV DNA是这种应用的基础。 具体目标是：1.开发血清学标记物来识别溶解性 循环蛋白; 2.分离感染性KSHV; 3.来定义 BCBL细胞中KSHV的潜伏性至裂解周期转换。的途径 开发血清学标记将是做免疫印迹， 使用BCBL细胞和患者来源的抗体的免疫荧光测定- 含有血清。一旦识别出与裂解周期相关的 蛋白质，将进行蛋白质的克隆和序列分析。 分离传染性病毒的方法最初将侧重于 BCBL线，然后进入患者源材料。靶细胞 已经测试的包括B和T细胞系，它们都被证明是阴性的。 对于这项拟议的工作，MRC 5细胞株，真皮的原代培养物， 将测试成纤维细胞和脐静脉内皮细胞， 将一些猴子肾细胞的细胞系。与BCBL线合作将 包括确定KSHV的大小和复杂性的实验 细胞系携带的基因组。它还将利用 观察KSHV能与EBV共感染B细胞 永生化细胞作为病毒分离的靶系统之一。 了解潜在的溶解周期转换的方法将是 以确定诱导物，然后定量群体中的细胞数量 以及定量和定性的 KSHV多肽和mRNA的产生。这种方法包括： 使用代表性差异分析和消减cDNA 图书馆.这将导致基因组和cDNA克隆的衍生 KSHV，用于定义即刻早期和早期KSHV BCBL系中的基因表达。这些工具与现有的 可用于EBV研究的工具，将有助于确定KSHV和 EB病毒在协调或不同的控制下。

DESCRIPTION (Adapted from investigator's abstract): This study is focused on the KSHV which has been found in KS and in biopsies of some non-Hodgkin's lymphoma. The BCBL line, which is latently infected with both KSHV and EBV, is the only known cell culture source for KSHV grown in cell culture, as well as the only human cells known to be latently infected with two human herpes viruses. Preliminary studies demonstrating that the BCBL line can be induced to express KSHV lytic cycle peptides and amplify KSHV DNA are the basis of this application. The specific aims are: 1. To develop serologic markers to identify lytic cycle proteins; 2. To isolate infectious KSHV; and 3. To define the latent to lytic cycle switch of KSHV in BCBL cells. The approach to develop serologic markers will be to do immunoblotting and immunofluorescence assays using BCBL cells and patient-derived antibody- containing sera. Upon recognition of the lytic cycle-associated proteins, cloning and sequence analyses of the proteins will be done. The approach towards isolating infectious virus will initially focus on the BCBL line, then proceed to patient source material. Target cells already tested included B and T cell lines, which all proved negative. For this proposed work, the MRC5 cell strain, primary cultures of dermal fibroblasts, and umbilical vein endothelial cells will be tested, as will some lines of monkey kidney cells. Work with the BCBL line will include experiments to determine the size and complexity of the KSHV genome carried by the cell line. It will also capitalize on the observation the KSHV can co-infect B cells with EBV, by using EBV immortalized cells as one of the target systems for virus isolation. The approach to understanding the latent to lytic cycle switch will be to identify inducers, then quantitate numbers of cells in populations that are induced and as well as the quantitative and qualitative production of KSHV polypeptides and mRNAs. Included in this approach is the use of representational difference analysis and subtraction cDNA libraries. This will lead to the derivation of genomic and cDNA clones of KSHV which would be used to define immediate early and early KSHV gene expression in the BCBL line. These tools, combined with existing tools available for the study of EBV, will help determine if KSHV and EBV are under coordinate or distinct control.