GAMMA INTERFERON RECEPTOR ON TUMORILYTIC MACROPHAGES

这项研究的长期目标是了解伽马射线是如何 干扰素受体参与巨噬细胞活化的调节 杀死肿瘤细胞 开展拟议的 研究表明，鼠γ干扰素（MuIFN-gamma）， 巨噬细胞活化因子，通过膜介导其作用 受体的 因此，对受体的研究应该产生 与识别早期事件相关的信息， 参与巨噬细胞活化的诱导。 的可用性 来自Genentech Inc.的大量重组鼠γ干扰素， 以及培养的巨噬细胞的同质群体使得这些 可行的研究。 它们将包括：（1）受体结合研究， 设计用于定量MuIFN-γ的数量和结合亲和力， 未受刺激的巨噬细胞上的受体，以及那些 对MuIFN-γ（炎性巨噬细胞）的反应性增加，或 降低的反应性（细胞后巨噬细胞）。 计算机辅助 Scatchard图的分析（LIGAND程序）将是主要方法 （2）从巨噬细胞富集MuIFN-γ受体复合物 膜，具有或不具有化学交联;（3）制备 针对受体的单克隆抗体。这些单克隆抗体 将用于免疫纯化受体，以及确定是否 抗受体抗体结合或不抗受体抗体结合可以在功能上模拟 MuIFN-γ;和（4）部分理化和生化 分离的MuIFN-γ受体的表征。 如果时间允许， 将进行额外的研究，以初步表征 受体的功能类型。 一般指征为 到功能类型，然后应该可以设计明确的 这些研究将展示受体如何启动诱导 activation. 这项拟议中的研究意义重大，因为它将产生 关于一种重要的宿主防御机制的新的基本信息， 巨噬细胞活化以杀死肿瘤细胞。 预计大部分 所获得的知识将同样适用于理解如何 MuIFN-γ调节多种其它巨噬细胞特征， 功能，从诱导抗病毒状态到吞噬细胞或 抗原呈递 据我们所知，这些将是第一次研究 巨噬细胞上的γ干扰素受体 （高频）

The long range goal of the proposed research is to learn how the gamma interferon receptor is involved in the regulation of macrophage activation for tumor cell killing. The rationale for undertaking the proposed research is that murine gamma interferon (MuIFN-gamma), which is one form of macrophage activating factor, mediates its effects through a membrane receptor. Investigation of the receptor should, therefore, yield information that is pertinent to identifying the early events that are involved in the induction of macrophage activation. The availability of large amounts of recombinant murine gamma interferon from Genentech Inc., as well as homogenous populations of cultured macrophages have made these studies feasible. They will include: (1) receptor binding studies, designed to quantify the number and binding affinity of MuIFN-gamma receptors on unstimulated macrophages, as well as those with either increased responsiveness to MuIFN-gamma (inflammatory macrophages) or diminished responsiveness (postcytolytlc macrophages). Computer-assisted analysis (LIGAND program) of Scatchard plots will be the principal approach here; (2) enrichment of the MuIFN-gamma receptor complex from macrophage membranes, either with or without chemical crosslinking; (3) preparation of monoclonal antibodies against the receptor. These monoclonal antibodies will be used to immunopurify the receptor, as well as to determine whether or not antireceptor antibody binding can functionally mimic binding of MuIFN-gamma; and (4) partial physicochemical and biochemical characterization of the isolated MuIFN-gamma receptor. If time allows, additional studies will be performed to preliminarily characterize the receptor with regard to its functional type. With general indications as to the functional type, it should then be possible to design definitive studies that will show how the receptor initiates the events that induce activation. The proposed research is significant because it will yield new, basic information about an important host defense mechanism, macrophage activation for tumor cell killing. It is expected that much of the knowledge obtained will be equally applicable to understanding how MuIFN-gamma modulates a variety of other macrophage characteristics and functions, ranging from induction of an antiviral state to phagocytes or antigen presentation. To our knowledge, these will be the first studies of the gamma interferon receptor on macrophages. (HF)