Biochemical studies and medical application of immunoregulatory factors

1. 人T细胞杂交瘤产生的淋巴因子：我们建立了一种利用代谢抑制剂（艾美汀和放线菌素D）构建人T细胞杂交瘤的新选择方法。从所建立的一个人T细胞杂交瘤克隆中，我们克隆了人淋巴蛋白的cDNA，并成功地产生了大量的重组人淋巴蛋白。在体外和体内均发现重组人淋巴毒素对恶性肿瘤细胞具有特异性的细胞毒作用。在分析人淋巴蛋白体内抗肿瘤活性的过程中，我们发现人淋巴蛋白具有强效的巨噬细胞趋化和活化巨噬细胞的活性。从另一个人T细胞杂杂瘤克隆H3-E9-6中，我们部分纯化了两种巨噬细胞激活因子。其中，MAF-CI是一种启动因子，与人γ - >-干扰素不同。另一种是MAF-CII，是一种不同于脂多糖的触发因子。这两种巨噬细胞活化因子对人单核细胞具有协同作用，诱导人单核细胞具有较强的杀瘤活性。目前我们正在对这两个因子进行完整的纯化和结构研究。人和小鼠巨噬细胞杂交瘤产生的单因子：我们成功地建立了许多小鼠和人巨噬细胞杂交瘤，并分析了这些巨噬细胞杂交瘤产生的各种单因子。其中一个已建立的人巨噬细胞杂交瘤克隆可分泌一种不同于人淋巴毒素、肿瘤坏死因子和白细胞介素-1的强效肿瘤特异性细胞毒素。

1. Lymphokines produced by human T cell hybridomas : We established a novel selection method for the construction of human T cell hybridomas using metabolic inhibitors (emetine and actinomycin D). From one of human T cell hybridoma clones thus established, we cloned cDNA for human lymphotoxin and succeeded to produce a large amounts of recombinant human lymphotoxin. The recombinant human lymphotoxin was found to exert potent cytotoxicity specifically against malignant cells in vitro and in vivo. On the process of the analysis of in vivo anti-tumor activity of human lymphotoxin, we found that human lymphotoxin has potent macrophage chemotactic and macrophage activating activities. From the other human T cell hybridoma clone, H3-E9-6, we partially purified two macrophage activating factors. One of them, MAF-CI, is a priming factor which is not identical with human <gamma>-interferone. The other, MAF-CII, is a triggering factor which is different from lipopolysaccharide. These two macrophage activating factors show a synergistic effect on human monocytes and induce strong tumoricidal activity of human monocytes. Now we are wroking on the complete purification and structural study of these two factors.2. Monokines produced by human and mouse macrophage hybridomas : We succeeded to establish a number of mouse and human macrophage hybridomas and analyzed various monokines produced by these macrophage hybridomas. One of the human macrophage hybridoma clones established was found to secrete a potent tumor-specific cytotoxin which was distinct from human lymphotoxin, tumor necrosis factor and interleukin-1.