IMMUNOREGULATORY EFFECTS OF THE INTERFERONS

干扰素介导多种生物学功能， 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; 我们假设它们是负生长调节剂 并与T细胞、B细胞和单核细胞相互作用， 免疫调节淋巴因子。 他们已经证明 在血液肿瘤和我们自己的肿瘤中具有显著的临床活性 数据暗示了直接的抗增殖作用机制 通过特定的细胞表面受体介导，可能作为第二个 导致癌基因调节的信息生长抑制剂。 尽管如此，它们并没有被广泛用作抗癌剂， 尽管存在干扰素的特异性受体， 各种肿瘤细胞系和新鲜切除的肿瘤。 到 进一步了解它们的分子生物学及其可能的作用 在淋巴因子网络中，我们现在建议制备特异性的 I型和II型干扰素受体的抗独特型抗体， 从Burkitt中纯化α-2干扰素的人受体 淋巴瘤细胞系Daudi，并探讨α和 γ干扰素和受体表达为阴性生长 干扰素敏感和抗性人细胞系中的调节剂 和肿瘤细胞。 重组α和γ干扰素将 被放射性标记并用于评估特异性受体结合， 周转和亚细胞定位。 我们将制备单克隆 小鼠中的抗体和兔中的多克隆抗体， I型和II型受体，或者使用纯化的受体 蛋白质和/或单克隆抗体的抗独特型的产生 我们实验室已经制备了抗体，可以结合 配体上的受体特异性结构域并阻断抗病毒和 抗增殖功能。 受体的纯化将是 使用亲和色谱法完成， 干扰素琼脂糖，抗受体抗体琼脂糖， 麦胚琼脂糖凝胶和HPLC。 氨基酸序列 纯化的受体和分离的胰蛋白酶肽将被 确定并用于构建几种寡核苷酸探针 用于筛选人类基因组DNA文库 包含在λ噬菌体中。 或者，抗受体 抗体或寡核苷酸探针可用于筛选cDNA 表达文库。杆菌 受体基因将被分离， 克隆，其核苷酸和侧翼序列确定为 以及染色体定位。 合适的基因片段 将用于分析正常和 癌症人体组织，以确定机制， 对干扰素作用敏感性和抵抗性及其作用 特定的致癌基因。 这些研究将阐明 干扰素系统，并可允许其临床应用于 与其他淋巴因子结合或可能识别 治疗人类肿瘤的替代治疗策略 恶性肿瘤

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.