TYPE D RETROVIRUS AND PULMONARY CARCINOMA

D 型逆转录病毒和肺癌

• 批准号: 3203168
• 负责人: James C DeMartini
• 金额: $ 23.9万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-03-01 至 1996-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3203168
• 关键词: Retroviridae; Retroviridae disease; diagnostic respiratory lavage; enzyme linked immunosorbent assay; gene expression; genetic transcription; humoral immunity; immunocytochemistry; immunodeficiency; in situ hybridization; lung neoplasms; neoplastic cell; northern blottings; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction; respiratory epithelium; sheep; southern blotting; viral carcinogenesis; virus RNA; virus infection mechanism; virus related neoplasm /cancer; virus replication; western blottings

Ovine pulmonary carcinoma (OPC) is a contagious lung tumor of sheep that originates from type II alveolar epithelial cells and strikingly resembles bronchioloalveolar carcinoma (BAC) of humans. Morphological, biochemical, and immunological data implicate a type D retrovirus as the cause of OPC, but this virus has not yet been propagated in cell culture. With the expectation that OPC will prove to be a valuable model of a retrovirus-induced pulmonary neoplasia that may lead to a greater understanding of pulmonary carcinogenesis we propose to: 1) Examine the distribution of the OPC virus within tumor cells to determine whether the viral generation is exogenous or endogenous, whether the virus is clonally integrated in the tumor cells, and to develop appropriate approaches for dissecting the mechanism of oncogenesis; and 2) Evaluate OPC viral product expression in infected animals prior to and after the onset of neoplastic sequelas. The experimental design is based on initially generating the appropriate reagents-molecular and immunological probes as well as tissues from animals from initial infection through the development of neoplastic sequelae. This will then be applied to correlate the viral life cycle (integration, expression and regulation) to the neoplastic processes. The methods to be used include: nucleic acid hybridizations, both from tissues (Southern and Northern blots, RNA dot blots) and in specific sites (in situ hybridization); and parallel immunological assays (ELISAs), immuno-histochemistry and Western blots). Reagents will be developed using the polymerase chain reaction for isolation, testing and cloning probes, and sequencing; generation of recombinant antigens will result from these clones and will be used along with purified viral components to generate OPC viral antisera. All of these techniques have been done by the investigators involved. Similar studies, in some cases without the advantage of the newer techniques, have been used with success in other studies of retroviral-associated neoplasia.

绵羊肺癌(OPC)是绵羊的一种传染性肺部肿瘤。 起源于II型肺泡上皮细胞，引人注目的是 类似于人类的细支气管肺泡癌(BAC)。形态上的， 生化和免疫学数据表明D型逆转录病毒是 引起OPC，但该病毒尚未在细胞培养中繁殖。 期望OPC将被证明是一种有价值的 逆转录病毒诱导的肺肿瘤可能导致更大的 对肺癌发生的理解，我们建议：1)检查 OPC病毒在肿瘤细胞内的分布以确定是否 病毒的产生是外源性的还是内源性的，无论病毒是 克隆性整合到肿瘤细胞中，并发展成适当的 肿瘤发生机制的研究方法；2)评价 OPC病毒产物在感染动物感染前后的表达 肿瘤后遗症的发作。实验设计的基础是 最初产生合适的试剂--分子和免疫学 探针以及动物的组织从最初的感染到 肿瘤后遗症的发展。然后这将应用于 与病毒生命周期(整合、表达和调控)相关 肿瘤的过程。将使用的方法包括：核 酸性杂交，来自组织(Southern和Northern blots，RNA 斑点杂交)和特定位置(原位杂交)；以及平行 免疫分析(ELISA)、免疫组织化学和Western blotts)。 将利用聚合酶链式反应开发试剂用于 分离、测试和克隆探针以及测序；产生 重组抗原将由这些克隆产生，并将用于 用纯化的病毒成分产生OPC病毒抗血清。所有的 这些技术都是由相关调查人员完成的。类似 在某些情况下，研究没有利用新技术的优势， 已经成功地用于其他逆转录病毒相关的研究 肿瘤。