EBNA AND OTHER VIRAL PRODUCTS IN EBV TRANSFORMED CELLS

本项目的目的是继续研究EBV- 编码的生长转化相关核(EBNA-1-6) 和膜(LMP)蛋白。的映射和定义 EBNA-4和EBNA-6将继续。关于EBNA-5的工作将 关注B95-8/Raji原型负线和 可能存在可供选择的特性。我们还打算 将EBNAs与EA/VCA在增殖期的定位进行比较 分化的上皮层。核内 将在EM层面研究EBNAs的分布，并 与其他核蛋白相比，如c-myc、N-myc、Cyclin 和sn-RNPs。EBNA家族的蛋白质将具有亲和力 在现有单克隆体的帮助下进行纯化。纯净的 将对蛋白质进行DNA依赖的ATPase测试，并 毒物异构酶I和II的活性。一种改进的麦凯试验将是 用于搜索EBNA-2上的特定DNA结合位点和 EBNA-5。EBNA-1、EBNA-2和LMP转基因小鼠肿瘤 这些品系将在同基因小鼠身上进行免疫原性测试。 相应的人类转染体将进行CTL检测- 体外敏感性、转化相关变化和新的 激活标记。表型研究将关注 对成套研究的EBNAs进行协调管理的可能性 EBV转换的子线，来自相同的EBV阴性BL 排队。转myc基因的人将进行“镜像实验” LCL已转变为更类似于BL型的表型。为了 将表型调节的研究扩展到其他细胞类型，a C3d/EBV(CR2)受体克隆将从 文库，并在各种EBV阴性细胞中表达。受体 阳性细胞会感染EBV并表达 转化与裂解周期相关的蛋白质将是 紧随其后。我们还将研究所有七种抗原的表达情况。 在鼻咽癌活检中。

The purpose of this project is to continue the study of the EBV- encoded, growth-transformation associated nuclear (EBNA-1-6) and membrane (LMP) proteins. The mapping and definition of EBNA-4 and EBNA-6 will be continued. The work on EBNA-5 will be focused on the B95-8/Raji prototype negative lines and the possible existence of alternative specificities. We also intend to localize the EBNAs vs EA/VCA in the proliferating, compared to differentiating layers of the epithelium. The intranuclear distribution of the EBNAs will be studied at the EM level and compared to other nuclear proteins, such as c-myc, N-myc, cyclin and sn-RNPs. Proteins of the EBNA family will be affinity purified with the help of the existing monoclonals. The purified proteins will be tested for DNA dependent ATPase, and to poisomerase I and II activities. A modified McKay assay will be used to search for specific DNA binding sites on EBNA-2 and EBNA-5. EBNA-1, EBNA-2 and LMP transfected murine tumor lines will be tested for immunogenicity in syngeneic mice. Corresponding human transfectants will be tested for CTL- sensitivity in vitro, transformation associated changes and new activation markers. Phenotypic studies will be concerned with possible coordinated regulation of the EBNAs studied in sets of EBV-converted sublines, derived from the same EBV negative BL line. "Mirror experiments" will be performed on myc-transfected LCLs that have shifted to a more BL-like phenotype. In order to extend the study of phenotypic regulation to other cell types, a C3d/EBV (CR2) receptor clones will be isolated from a cDNA library and expressed in various EBV negative cells. Receptor positive cells will be infected with EBV and the expression of the transformation vs. lytic cycle associated proteins will be followed. We shall also study the expression of all seven antigens in nasopharyngeal carcinoma biopsies.