IMMUNOREGULATORY EFFECTS OF THE INTERFERONS

干扰素调节多种生物功能， 包括诱导分化、抑制正常和 肿瘤细胞生长，并逆转几种癌基因的作用； 我们假设它们起负增长调节器的作用 并与T细胞、B细胞和单核细胞相互作用 免疫调节性淋巴因子。他们已经证明了 血液肿瘤和我们自身的显著临床活动性 数据暗示了一种直接的、抗增殖的作用机制 通过特定的细胞表面受体，可能作为第二个 导致癌基因调控的消息生长抑制因子。 然而，它们并没有被广泛用作抗癌药物。 尽管存在干扰素的特异性受体 各种肿瘤细胞系和新移植的肿瘤。至 进一步了解它们的分子生物学和可能的作用 在淋巴因子网络中，我们现在建议准备特定的 I型和II型干扰素受体的抗独特型抗体 从Burkitt分离纯化人α-2干扰素受体 淋巴瘤细胞系Daudi，并探讨其在细胞周期中的作用。 干扰素及其受体表达呈负生长状态 干扰素敏感和耐药人类细胞系中的调节因子 和肿瘤细胞。重组α和伽马干扰素将 被放射性标记并用于评估特定受体结合， 周转和亚细胞定位。我们将准备单克隆体 小鼠的抗体和兔的多克隆抗体 I型和II型受体交替使用纯化的受体 蛋白质和/或产生抗独特型抗体 我们实验室已经制备的与之结合的抗体 配体上的受体特异性结构域并阻断抗病毒和 抗增殖作用。受体的纯化将是 使用亲和层析技术完成 干扰素琼脂糖基上抗受体抗体琼脂糖基 小麦胚芽琼脂糖凝胶，并用高效液相色谱法。氨基酸序列 纯化的受体和分离的胰蛋白酶多肽的 确定并用于构建几种寡核苷酸探针 将用于筛选人类基因组DNA文库 包含在Lambda噬菌体中。或者，抗受体 抗体或寡核苷酸探针可用于筛选cDNA 在大肠杆菌中构建表达文库。受体基因将被分离， 克隆，其核苷酸和侧翼序列被确定为 以及它的染色体定位。合适的基因片段 将被用来分析正常和 以确定人体癌变组织的作用机制 干扰素的作用及其影响的敏感性和耐药性 关于特定的致癌基因。这些研究将阐明 干扰素系统，并可能允许其临床应用于 与其他淋巴因子结合或可能识别 人类多发性硬化症治疗的替代治疗策略 恶毒。

The interferons mediate a wide variety of biologic functions, including the ability to induce differentiation, inhibit normal and tumor cell growth, and reverse the action of several oncogenes; we hypothesize that they function as negative growth regulators and interact with T cells, B cells and monocytes as immunoregulatory lymphokines. They have demonstrated significant clinical activity in hematologic neoplasms and our own data imply a direct, antiproliferative mechanism of action mediated via specific cell surface receptors, possibly as a second message growth inhibitor leading to oncogene modulation. Nevertheless, they are not widely used as anti-cancer agents in spite of the presence of specific receptors for interferon on a variety of tumor cell lines and freshly explanted tumors. To understand further their molecular biology and their possible role in the lymphokine network, we now propose to prepare specific anti-idiotype antibodies to type I and II interferon receptors, to purify the human receptor for alpha-2 interferon from the Burkitt lymphoma cell line Daudi, and to explore the role of alpha and gamma interferon and receptor expression as negative growth regulators in interferon sensitive and resistant human cell lines and tumor cells. Recombinant alpha and gamma interferon will be radiolabeled and used to assess specific receptor binding, turnover and subcellular localization. We will prepare monoclonal antibodies in mice and polyclonal antibodies in rabbits specific for the type I and II receptor using, alternatively, purified receptor proteins and/or the production of anti-idiotypes to monoclonal antibodies already prepared in our laboratory that bind to receptor-specific domains on the ligands and block antiviral and anti-proliferative function. Purification of the receptor will be accomplished using affinity chromatography on interferonsepharose, on anti-receptor antibody sepharose, on wheat germ sepharose, and with HPLC. The amino acid sequence of the purified receptor and of isolated tryptic peptides will be determined and used to construct several oligonucleotide probes that will be used to screen a genomic human DNA library contained in lambda phage. Alternatively, the anti-receptor antibodies or oligonucleotide probes can be used to screen a cDNA expression library in E. coli. The receptor gene will be isolated, cloned, and its nucleotide and flanking sequences determined as well as its chromosome localization. Appropriate gene fragments will be used to analyze the DNA and RNA in normal and cancerous human tissues to determine the mechanism of susceptibility and resistance to interferon's action and its effects on specific oncogenes. These studies will clarify the biology of the interferon system and may permit its clinical use in combination with other lymphokines or possibly identify alternative therapeutic strategies in the treatment of human malignancy.