AIDS LYMPHOMA HARBORING TWO GAMMA HERPESVIRUSES

描述(改编自调查人员摘要)：这项研究是 重点关注在KS和一些组织活检中发现的KSHV 非霍奇金淋巴瘤。BCBL线，它潜伏着感染 KSHV和EBV都是已知的KSHV培养源 在细胞培养中，以及已知的唯一潜伏的人类细胞 感染了两种人类疱疹病毒。初步研究 证明BCBL细胞可被诱导表达KSHV裂解物 环肽和扩增KSHV DNA是这一应用的基础。 其具体目的是：1.建立鉴定溶血的血清学标记物 环状蛋白；2.分离感染性KSHV；3.确定 KSHV在BCBL细胞中潜伏的裂解周期转换。接近的方法 开发血清学标记将进行免疫印迹和 使用BCBL细胞和患者来源的抗体进行免疫荧光分析- 含有血清的。在认识到与裂解周期相关的 将对蛋白质进行克隆和序列分析。 分离传染性病毒的方法最初将重点放在 BCBL行，然后继续患者来源材料。目标单元格 已经测试的包括B和T细胞系，结果都是阴性的。 在这项拟议的工作中，MRC5细胞株，真皮的原代培养 成纤维细胞和脐静脉内皮细胞将被检测为 会不会有一些猴子肾脏细胞。与BCBL线路合作将 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 用EBV观察KSHV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。 理解潜伏到溶解周期转换的方法将是 确定诱导剂，然后量化种群中的细胞数量 是归纳的，以及定量和定性的 KSHV多肽和mRNAs的生产。此方法包括 代表性差异分析和差减cDNA的应用 图书馆。这将导致基因组和cdna克隆的衍生。 将被用来定义即时早期和早期KSHV 基因在BCBL系中的表达。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 EBV处于协调或不同的控制之下。

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.