EPSTEIN BARR VIRUS EXPRESSION IN NORMAL HUMAN EPITHELIUM

EB病毒原代细胞类型的鉴定 (EBV)复制对于了解病毒的生物学是必不可少的， 传播方式和发病机制。我们对EBV的了解几乎都是 来自对与淋巴组织有关的病毒的研究。然而，EBV 与三种完全不同的疾病有关，不仅涉及 淋巴样成分，也包括上皮成分。 这项研究的目标是确定细胞分化对 EB病毒与上皮细胞的相互作用。膜受体的差异 表达，EBV特异性抗原表达，转录模式 EBV基因组和病毒生产力将在这两个地方进行检查 未分化和已分化的上皮细胞亚群。这些 将通过间接免疫荧光技术进行检测； 电子显微镜；DNA-RNA细胞杂交与新一代 克隆的生物素化的DNA探针；通过标准的淋巴细胞转化 化验。宿主细胞控制感染的研究现在才有可能 随着这种上皮细胞系统技术的建立 无分化外植体原代培养的长期保存 细胞分化的状态和同步诱导。 宿主细胞明显缺乏对EB病毒致病机制的影响 受体也将通过感染和共培养来解决 自体淋巴样细胞和上皮细胞群。证据： EBV在上皮细胞中的转化潜力将在 野生型EBV株感染或转基因的长期细胞培养 使用克隆的EBV DNA片段或来自自体的细胞DNA 淋巴母细胞培养。细胞易于通过，能够在 软琼脂和裸鼠体内的致瘤性将被评估。 良性肿瘤的细胞杂交分析将寻找临床相关性 腮腺和唾液腺的淋巴上皮病变以及 中国EBV基因组选择性编码区的划分 不同病例组的口咽上皮细胞。

Identification of the primary cell type in which the Epstein-Barr virus (EBV) replicates is essential to understanding the biology of the virus, mode of transmission and pathogenesis. Virtually all we know about EBV comes froms studies of the virus in relation to lymphoid tissue. Yet, EBV is linked to three quite different diseases with involvement of not only lymphoid but also epithelial elements. The goal of this study is to define the effects of cell differentiation on EBV-epithelial cell interaction. Differences in membrane receptor expression, EBV-specific antigen expression, transcriptional patterns of the EBV genome, and virus productivity will be examined in both undifferentiated and differentiated epithelial cell subsets. These determinations will be made by indirect immunofluorescence techniques; electron microscopy; DNA-RNA cytohybridization with a new generation of cloned, biotinylated DNA probes; and by standard lymphocyte transformation assays. Studies of host cell control of infection are only now possible in this epithelial cell system with the establishment of techniques for long-term maintenance of primary explant cultures in an undifferentiated state and synchronized induction of cell differentiation. The problems imposed on EBV pathogenesis by the apparent lack of host cell receptors will also be addressed by infection and cocultivation of autologous lymphoid and epithelial cell populations. Evidence for transforming potential of EBV in epithelial cells will be sought in long-term cell cultures infected with wild-type EBV strains or transfected with either cloned EBV DNA fragments or cellular DNA from autologous lymphoblastoid cell cultures. Ease of cell passage, ability to grow in soft agar, and oncogenicity in nude mice will be assessed. Clinical correlates will be sought by cytohybridization analyses of benign lymphoepithelial lesions of the parotid and salivary glands as well as by delineation of selectively encoded regions of the EBV genome in oropharyngeal epithelial cells from various patient groups.