EBNA AND OTHER VIRAL PRODUCTS IN EBV TRANSFORMED CELLS

本项目主要研究EB病毒的三个编码区 (EBV)在病毒转化中有规律地转录的基因组，但是 病毒非生产细胞。 这些区域被指定为LT-1， LT-2和LT-3区域。 我们打算：i）确定这些区域的基因产物并绘制它们的图谱 最近测序的EB病毒基因组。 ii）纯化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：该区域是否编码ACIF β核抗原， 在蛋白质印迹上被鉴定为82 K蛋白质 它是否负责 启动B淋巴细胞转化，包括多克隆活化？ 对于Lt-2：已知该区域编码EBNA-1，主要的 EBV转化细胞中的核抗原。 我们最近成功地生产了 用合成的抗EBNA抗体免疫家兔， 从LT-2的IR 3亚区推导的肽正在被开发用于 EBNA-1的亲和纯化。 对于LT-3：这个区域是否编码我们所知的EB病毒特异性抗原 最近通过测试来自EBV转化的病毒的膜组分检测到， 白细胞迁移抑制（LMI）试验中的非生产细胞？ 是 这种膜抗原与EBV诱导的膜抗原LYDMA相关， 免疫监视的主要目标 对于所有三个区域： 每种蛋白质产品单独地和以各种组合， 转染实验。 用小鼠和大鼠进行 成纤维细胞，也与正常或肿瘤的人B淋巴细胞 起源 我们最近在这一奋进中特别受到鼓舞， 用电击法成功地分离出淋巴细胞。

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.