LYMPHOCYTE/TUMOR INTERACTIONS IN SCID/HU CHIMERIC MICE

我们建议开发一种流行的人类癌症的小动物模型。 我们认为将适合对该建筑群的研究 肿瘤进展与免疫反应的相互关系。为 这些初步研究我们的工作将集中在卵巢腺癌 由于其医学意义，可获得适当的 标本，以及这种疾病的解剖学局限性质。这个 这种模式的发展取决于三个要素：1)我们的能力 将人恶性卵巢腹水分离成高度浓缩的种群 逆流离心法提取肿瘤细胞、淋巴细胞和单核细胞 洗脱(CCE)；2)我们建立人类的经验 严重联合免疫缺陷(SCID)小鼠的异种淋巴移植；以及 3)其他实验室发表的研究表明，人类卵巢 可以在常规的nu/nu小鼠身上建立肿瘤异种移植。 卵巢腹水细胞大量可用作为一种副产品 治疗性穿刺术。我们能够处理多达2x10-10个细胞 一次淋洗，提供了足够数量的活肿瘤细胞， 即使它们占腹水的比例相对较小，或者 与低总体生存能力的渗出物混合在一起。因为这件事 肿瘤主要局限于腹膜，尤其是 适合于异种移植研究；人类肿瘤和人类 淋巴细胞/单核细胞群可以在腹膜内建立 SCID小鼠的空洞。本提案的范围为：1) 纯化的人肿瘤细胞在SCID小鼠体内的建立；2) 肿瘤联合肿瘤浸润性接种的效果评价 淋巴细胞、单核细胞和外周血单核细胞； 提供肿瘤体外相关因素的初步表征 用肿瘤浸润性淋巴细胞和 单核细胞。通过这样做，我们希望提供一个系统，使我们能够 测试有关肿瘤逃逸潜在机制的假说 免疫根除，特别是在效应细胞层面 一代。虽然目前的提案侧重于卵巢癌，但我们 我相信这一方法将适用于其他肿瘤，因为 井。分离肿瘤浸润性炎症细胞的能力 高数量和高纯度使该系统特别适合于 未来对肿瘤特异性免疫的研究。

We propose to develop a small animal model of a prevalent human cancer that we believe will be suitable for the study of the complex interrelationship between tumor progression and the immune response. For these initial studies our work will focus on ovarian adenocarcinoma because of its medical significance, the availability of appropriate specimens, and the anatomically confined nature of the disease. The development of this model depends upon three elements: 1) our ability to separate human malignant ovarian ascites into highly enriched populations of tumor cells, lymphocytes, and monocytes by counterflow centrifugal elutriation (CCE); 2) Our experience with the establishment of human lymphoid xenografts in severe combined immunodeficient (SCID) mice; and 3) The published work of other laboratories indicating that human ovarian neoplastic xenografts can be established in conventional nu/nu mice. Ovarian ascites cells are available in large number as a byproduct of therapeutic paracentesis. We are able to process up to 2xl0-10 cells in a single elutriation run, providing ample numbers of viable tumor cells, even when they comprise a relatively small proportion of the ascites or are admixed with infiltrates of low overall viability. Because this tumor is primarily confined to the peritoneum, it is particularly well suited for xenografting studies; both human tumors and human lymphocyte/monocyte populations can be established within the peritoneal cavities of SCID mice. The scope of the present proposal is to: 1) Optimize the establishment of purified human tumor cells in SCID mice; 2) Assess the effect of co-inoculation of tumor plus tumor infiltrating lymphocytes, monocytes and peripheral blood mononuclear cells; and 3) Provide a preliminary characterization of in vitro correlates of tumor specific immunity as measured using tumor infiltrating lymphocytes and monocytes. In doing so we hope to provide a system that will allow us to test hypotheses concerning potential mechanisms by which tumor evades immune eradication, particularly at the level of effector cell generation. Although the present proposal focuses on ovarian cancer, we believe that this methodology will be applicable to other neoplasms as well. The ability to isolate tumor infiltrating inflammatory cells in high number and purity makes this system particularly well suited to future studies of tumor specific immunity.