TYPE D RETROVIRUS AND PULMONARY CARCINOMA

绵羊肺癌（OPC）是绵羊的传染性肺肿瘤， 来源于II型肺泡上皮细胞， 类似于人类的细支气管肺泡癌（BAC）。 形态学上， 生物化学和免疫学数据表明，D型逆转录病毒是 OPC的原因，但这种病毒尚未在细胞培养中繁殖。 期望OPC将被证明是一个有价值的模型， 逆转录病毒引起的肺肿瘤，可能导致更大的 了解肺癌的发生，我们建议：1）检查 OPC病毒在肿瘤细胞内的分布，以确定 病毒的产生是外源性的还是内源性的，无论病毒是 克隆整合在肿瘤细胞中，并发展适当的 探讨肿瘤发生机制的方法; 2）评估 感染动物中的OPC病毒产物表达之前和之后， 肿瘤后遗症的发作。 实验设计基于 最初产生适当的试剂-分子和免疫试剂 探针以及来自动物的组织，从最初的感染到 肿瘤后遗症的发展。 这将适用于 关联病毒生命周期（整合、表达和调控） 肿瘤的过程。 使用的方法包括：核酸 酸杂交，均来自组织（Southern和北方印迹，RNA 斑点杂交）和特异性位点（原位杂交）;以及 免疫学测定（ELISA）、免疫组织化学和蛋白质印迹）。 将使用聚合酶链反应开发试剂， 分离、测试和克隆探针，以及测序; 重组抗原将由这些克隆产生，并将沿着 与纯化的病毒组分反应以产生OPC病毒抗血清。 所有 这些技术是由有关的调查人员采用的。 类似 研究，在某些情况下，没有新技术的优势， 已成功用于其他逆转录病毒相关的研究， 肿瘤形成

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.