VIRAL ONCOGENES, INTERFERON, AND IMMUNITY

病毒癌基因、干扰素和免疫

• 批准号: 2067045
• 负责人: John Michael Routes
• 金额: $ 9.79万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2067045
• 关键词: Adenoviridae; cell cycle proteins; cellular immunity; chemical binding; gene mutation; genetic mapping; genetic regulation; human papillomavirus; human tissue; interferons; natural killer cells; oncogenes; oncoproteins; simian virus 40; tissue /cell culture; transfection; virus genetics; Adenoviridae; cell cycle proteins; cellular immunity; chemical binding; gene mutation; genetic mapping; genetic regulation; human papillomavirus; human tissue; interferons; natural killer cells; oncogenes; oncoproteins; simian virus 40; tissue /cell culture; transfection; virus genetics

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.

天然杀手（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（IFN） 通过激活促进AD2/5感染细胞的选择性裂解 NK细胞并保护未感染的细胞（IFN介导 细胞保护或IFN MCP），但未来自NK细胞的AD感染细胞 介导的裂解。 单个AD基因的表达，早期区域1a （EIA），阻止IFN MCP。 这提供了第一个学习的机会 单个病毒基因的IFN MCP干扰。 初步的 遗传分析表明，序列在第一个中的表达 EIA外显子抑制IFN MCP是必要的。 表达 SV40 LT，与第一个外显子共享同源序列 EIA，也抑制IFN MCP。 EIA和SV40的同源区域 LT最近已显示与特定的细胞蛋白结合 （例如，P105视网膜细胞瘤（RB）基因产物； P107； P60，细胞周期蛋白A） 与其中的一些生物学活动相关 病毒癌蛋白。 基于这些研究，看来 SV40 LT和AD EIA对IFN MCP的抑制是通过 相同的细胞途径，可能是通过与特定的，常见的相互作用 细胞蛋白（S）。 该提议的第一个具体目的：通过突变来映射 分析，负责ELA抑制的遗传区域 IFN MCP并确定突变EIA蛋白的能力是否 结合特定的细胞蛋白（p60（Cyclin A），P105（RB），PL07， p300）与ELA介导的IFN MCP抑制相关。 人类乳头瘤病毒（HPV）早期7（E7），例如SV40 LT，共享 具有ELA的第一个外显子的同源序列。然而， 初步研究表明，HeLa细胞中的HPV E7表达为 不是阻止IFN MCP。 该提案的第二个具体目的：确定表达是否表达 其他类型的人类细胞中的HPV16或18 E7基因产物 抑制IFN MCP。 将使用以下方法：a） SV40 LT或HPV16 E7中IFN MCP的比较表达角质形成细胞 和成纤维细胞细胞系。 b）确定重要性 IFN诱导的HPV16/18中E7表达的下调转换 人体细胞系调节IFN MCP的抑制作用。 c） 确定E7表达水平与 IFN MCP。