EBNA AND OTHER VIRAL PRODUCTS IN EBV TRANSFORMED CELLS

本项目重点研究了EB病毒的三个编码区 (EBV)在病毒转化中定期转录的基因组，但 病毒非产生细胞。这些区域被指定为LT-1， LT-2和LT-3区。 我们打算：i)识别这些区域的基因产物，并绘制它们的图谱 到最近测序的EBV基因组。 2)提纯基因产物并研究其性质。 3)研究单个基因的生物学功能。 对于LT-1：这个区域是否编码ACIF可染色的核抗原， 在蛋白质印迹中被鉴定为82K蛋白？它对这起事件负责吗 启动B淋巴细胞转化，包括多克隆激活？ 对于LT-2：该区域已经知道EBNA-1的编码，主要 EBV转化细胞中的核抗原。我们最近在生产方面取得的成功 人工合成EBNA抗体免疫兔的研究 从LT-2的IR3亚区衍生的多肽正在被开发用于 EBNA-1的亲和纯化。 对于LT-3：这个区域编码的是我们已有的EBV特异性抗原吗 最近通过检测EBV转化的病毒的膜组分检测到的 白细胞迁移抑制(LMI)试验中的非生成性细胞？是 这种膜抗原与EBV诱导的膜抗原LYDMA有关， 免疫监测的主要目标是什么？ 对于所有这三个区域：我们希望研究 每种蛋白质产品分别，并以不同的组合，通过 转基因实验。它们是用老鼠和大鼠进行的 成纤维细胞，以及正常或肿瘤的人B淋巴细胞 起源。我们在这方面的努力尤其受到鼓舞，因为我们最近 电击法成功转导淋巴样细胞。

This project focuses on the three coding regions of the Epstein-Barr virus (EBV) genome that are regularly transcribed in virally transformed, but virus non-producer cells. These regions have been designated as the LT-1, LT-2 and LT-3 regions. We intend to: i) Identify the gene products of these regions and map them to the recently sequenced EBV genome. ii) Purify the gene products and study their properties. iii) study the biological functions of the individual genes. For LT-1: Does this region code for an ACIF stainable nuclear antigen, identified as an 82K protein on Western blots? Is it responsible for the initiation of B-lymphocyte transformation, including polyclonal activation? For Lt-2: This region is already known to code for EBNA-1, the main nuclear antigen in EBV-transformed cells. Our recent success in producing EBNA-specific antibodies in rabbits by immunizing them with synthetic peptides deduced from the IR3 subregion of LT-2 is being exploited for the affinity purification of EBNA-1. For LT-3: Is this region coding for the EBV-specific antigen that we have recently detected by testing membrane fractions from EBV-transformed, virus non-producer cells in the leukocyte migration inhibition (LMI) test? Is this membrane antigen related to the EBV-induced membrane-antigen, LYDMA, the main target for immune surveillance? For all three regions: We wish to study the functional significance of each protein product separately, and in various combinations, by transfection experiments. They are carried out with mouse and rat fibroblasts, and also with human B-lymphocytes of normal or neoplastic origin. We are particularly encouraged in this endeavor by our recent success in transfecting lymphoid cells by using electroshock method.