CYTOKINE AND ANTI-RECEPTOR ANTIBODY FUSION PROTEINS

主要目标是确定能够实质性地 降低自发转移中肿瘤细胞的转移率 人类和小鼠黑色素瘤的模型。需要检验的中心假说 新的免疫疗法设计的目的是激活免疫 效应细胞或破坏肿瘤血管是足够有效的 来实现这一目标。为实现这些目标而设计的具体目标 目的包括测定重组人的体外活性 重组人白细胞介素2、重组肿瘤坏死因子与嵌合体融合蛋白的构建 人/鼠抗人表皮生长因子-1的单抗 受体，这些涉及：a)抗原结合；b)功能性细胞因子 活性；以及c)免疫效应器激活肿瘤细胞裂解 细胞。单抗-细胞因子融合蛋白的体内参数 包括它们在小鼠体内的清除率和它们的 自发转移性BL6黑色素瘤小鼠体内的生物分布。 我们将调整我们已建立的同基因转移模型，使肿瘤 细胞表达与人类相同的分子定义的靶抗原 构成我们黑色素瘤基础的黑色素瘤细胞系M24met和C8161 SCID小鼠肿瘤转移模型的建立。因此，B16黑色素瘤细胞将稳定 将编码人表皮生长因子-γ的c DNA导入细胞内。我们的药效 抗体-细胞因子融合蛋白降低肿瘤细胞生长速度的研究 在我们的同基因黑色素瘤转移模型中 与单抗、细胞因子或其混合物所达到的效果相比 两者都有。一旦mAb-细胞因子融合蛋白被证明是最有效的 在这方面，这一进程所涉及的假定机制将是 检查过了。这将包括通过mAb-IL-2裂变激活T细胞 蛋白质与单抗-肿瘤坏死因子融合对肿瘤血管的破坏 蛋白质。一旦融合蛋白的疗效及其机制 在完全免疫活性的小鼠黑色素瘤中已经确定了作用 转移模型，将努力建立这些参数在 我们的人黑色素瘤在SCID小鼠体内的转移模型。这将涉及到 利用选定和定义的人类种群进行选择性重建 淋巴效应细胞和肿瘤血管破坏的评估。 预计这些研究的一些结果可能有助于 转移性肿瘤治疗新方法的发展 黑色素瘤。

The major objective is to identify approaches capable of substantially reducing the rate of tumor cell dissemination in spontaneous metastasis models of human and murine melanoma. The central hypothesis to be tested is that novel immunotherapy modalities designed either to activate immune effector cells or to destroy tumor vasculature are sufficiently effective to achieve this objective. Specific aims designed to accomplish these objectives include a determination of in vitro activities of recombinant fusion proteins constructed between rlL-2 and rTNF and chimeric human/mouse monoclonal antibodies (mAb) directed against human EGF- receptor, These involve: a) antigen binding; b) functional cytokine activities; and c) activation of tumor cell lysis by immune effector cells. In vivo parameters of the mAb-cytokine fusion proteins that will be evaluated include their clearance rates in mice and their biodistribution in mice bearing spontaneously metastasizing Bl6 melanoma. We will adapt our established syngeneic metastatic model so that the tumor cells express the same molecularly-defined target antigens as the human melanoma cell lines M24met and C8161 that form the basis for our melanoma metastasis models in SCID mice. Thus, B16 melanoma cells will be stably transfected with cDNA encoding for human EGF-r. The efficacy of our antibody-cytokine fusion proteins to reduce the rate of tumor cell growth and dissemination in our syngeneic melanoma metastasis model will be compared with that achieved by either mAbs, cytokines, or a mixture of both. Once mAb-cytokine fusion proteins are shown to be most effective in this regard, putative mechanisms involved in this process will be examined. These will include T-cell activation by mAb-IL-2 fission proteins and destruction of the tumor vasculature by mAb-TNF fusion proteins. Once the efficacy of the fusion proteins and their mechanisms of action have been established in the fully immunocompetent murine melanoma metastasis model, efforts will be made to establish these parameters in our human melanoma metastasis model in SCID mice. This will involve selective reconstitution with selected and defined populations of human lymphoid effector cells and evaluations of tumor vasculature destruction. It is anticipated that some of the results of these studies may contribute toward the development of novel approaches for the treatment of metastatic melanoma.