VIRAL ONCOGENES, INTERFERON, AND IMMUNITY

自然杀伤（NK）细胞是细胞免疫的重要组成部分。 抗病毒免疫 最近的研究表明，干扰素（IFN） 通过激活Ad 2/5，促进Ad 2/5感染细胞的选择性裂解 NK细胞和保护未感染的细胞（IFN介导的 细胞保护或IFN MCP），而不是Ad感染的细胞从NK细胞 介导的裂解。 单个Ad基因，早期区域1A的表达 (EIA)，阻断IFN MCP。 这是第一次有机会学习 单个病毒基因对IFN MCP的干扰作用。 初步 遗传分析表明，表达的序列在第一个 EIA的外显子是抑制IFN MCP所必需的。 表达 SV 40 LT与SV 40的第一个外显子具有同源序列， EIA也抑制IFN MCP。 EIA和SV 40的同源区域 LT最近被证明可以结合特定的细胞蛋白质 (e.g., p105视网膜母细胞瘤（Rb）基因产物; p107; p60，细胞周期蛋白A） 这与它们的一些生物活性有关， 病毒癌蛋白 根据这些研究， SV 40 LT和Ad EIA对IFN MCP的抑制作用是通过 相同的细胞途径，可能通过与特定的，共同的 细胞蛋白。 这项建议的第一个具体目标：通过突变 分析，负责ElAs抑制的遗传亚区 并确定突变EIA蛋白是否能够 结合特定的细胞蛋白（p60（细胞周期蛋白A），p105（Rb），p107， p300）与ElA介导的IFN MCP抑制相关。 人乳头瘤病毒（HPV）早期区域7（E7），如SV 40 LT，共享 与ElA第一外显子同源的序列。然而，在这方面， 初步研究表明，HPV E7在HeLa细胞中的表达确实 不阻断IFN MCP。 本建议的第二个具体目标：确定表达 HPV 16或18 E7基因产物在其他类型的人类细胞中 抑制IFN MCP。 将采用以下方法： IFN MCP在SV 40 LT和HPV 16 E7表达的角质形成细胞中的比较 和成纤维细胞系。B）确定以下各项的显著性： IFN诱导的HPV 16/18转化细胞E7表达下调 人细胞系调节IFN MCP的抑制。c）、 确定E7表达水平与细胞凋亡之间的关系。 IFN MCP。

Natural killer (NK) cells are an important component of cellular antiviral immunity. Recent studies show that interferon (IFN) promotes the selective lysis of Ad2/5 infected cells by activating NK cells and protecting uninfected cells (IFN mediated cytoprotection or IFN MCP) but not Ad infected cells from NK cell mediated lysis. Expression of a single Ad gene, early region 1A (EIA), blocks IFN MCP. This offers the first opportunity to study the interference of IFN MCP by a single viral gene. Preliminary genetic analysis reveals that expression of sequences in the first exon of EIA is necessary for inhibition of IFN MCP. Expression of SV40 LT, which shares homologous sequences with the first exon of EIA, also inhibits IFN MCP. The homologous regions of EIA and SV40 LT have been recently shown to bind specific cellular proteins (e.g., p105 retinoblastoma (Rb) gene product; p107; p60, cyclin A) which has con-elated with some of the biological activities of these viral oncoproteins. Based on these studies, it appears that the inhibition of IFN MCP by SV40 LT and Ad EIA is mediated through the same cellular pathway, likely by interacting with specific, common cellular protein(s). The first specific aim of this proposal: To map, by mutational analysis, the genetic subregion(s) responsible for ElAs inhibition of IFN MCP and to determine if the ability of mutant EIA proteins to bind specific cellular proteins (p60 (cyclin A), p105 (Rb), pl07, p300) correlates with ElA mediated inhibition of IFN MCP. Human papillomavirus (HPV) early region 7 (E7), like SV40 LT, share homologous sequences with the first exon of ElA . However, preliminary studies show that HPV E7 expression in HeLa cells does not block IFN MCP. The second specific aim of this proposal: To determine if expression of HPV16 or 18 E7 gene products in other types of human cells inhibits IFN MCP. The following approaches will be used: a) Comparison of IFN MCP in SV40 LT or HPV16 E7 expressing keratinocyte and fibroblast cell lines. b) Determination of the significance of IFN-induced, downregulation of E7 expression in HPV16/18 transformed human cell lines in modulating the inhibition of IFN MCP. c) Determination of the relationship between level of E7 expression and IFN MCP.