VIRAL ONCOGENES, INTERFERON, AND IMMUNITY

自然杀伤(NK)细胞是细胞的重要组成部分 抗病毒免疫。最近的研究表明干扰素(干扰素) 通过激活促进AD2/5感染细胞的选择性溶解 NK细胞与保护未感染细胞(干扰素介导) 细胞保护或干扰素MCP)，但不是来自NK细胞的Ad感染细胞 中介裂解。单个Ad基因早期区域1A的表达 (EIA)，阻断干扰素单核细胞趋化蛋白。这提供了第一次学习的机会 单个病毒基因对干扰素MCP的干扰。初步 遗传分析表明，第一个序列的表达 EIA外显子是抑制干扰素单核细胞趋化蛋白的必要条件。的表达 SV40 LT，与V40的第一个外显子具有同源序列 EIA，也抑制干扰素单核细胞趋化蛋白。EIA和SV40的同源区 最近发现它可以结合特定的细胞蛋白。 (例如，p105视网膜母细胞瘤(RB)基因产物；p107；p60，细胞周期蛋白A) 这与这些化合物的一些生物活性有关 病毒癌蛋白。根据这些研究，似乎 SV40 LT和Ad EIA对干扰素MCP的抑制作用是通过 相同的细胞途径，很可能是通过与特定的、常见的 细胞蛋白(S)。 这项提议的第一个具体目标是：通过突变绘制地图 分析，导致ELA抑制的遗传亚区(S) 并确定突变的EIA蛋白是否具有 结合特定的细胞蛋白(p60(细胞周期蛋白A)、p105(Rb)、pl07、 P300)与ELA介导的干扰素单核细胞趋化蛋白抑制相关。 人乳头瘤病毒(HPV)早期第7区(E7)与SV40 LT一样，分享 与ELA第一外显子同源的序列。然而， 初步研究表明，HPV E7在HeLa细胞中的表达 不阻断干扰素单核细胞趋化蛋白。 这项提议的第二个具体目标是：确定是否表达 HPV16或18 E7基因产物在其他类型的人类细胞中的表达 抑制干扰素单核细胞趋化蛋白。将使用以下方法：a) 干扰素-单核细胞趋化蛋白在表达SV40 LT和HPV16 E7角质形成细胞中的比较 和成纤维细胞系。B)确定以下方面的重要性 干扰素诱导HPV16/18转化细胞E7表达下调 人细胞株对干扰素单核细胞趋化蛋白的抑制作用。c) E7基因表达水平与E7基因表达的关系 干扰素单核细胞趋化蛋白。

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.