CYTOKINE AND ANTI-RECEPTOR ANTIBODY FUSION PROTEINS

主要目标是确定能够实质性地 降低自发转移中的肿瘤细胞播散率 人和鼠黑素瘤模型。待检验的中心假设 是新的免疫治疗方式， 效应细胞或破坏肿瘤脉管系统是足够有效的 以实现这一目标。旨在实现这些目标的具体目标 目的包括测定重组的体外活性， 在rIL-2和rTNF之间构建的融合蛋白和嵌合 抗人EGF的人/小鼠单克隆抗体（mAb） 这些包括：a）抗原结合; B）功能性细胞因子 活性;和c）通过免疫效应物激活肿瘤细胞溶解 细胞 mAb-细胞因子融合蛋白的体内参数， 包括它们在小鼠中的清除率和它们的 在携带自发转移的B16黑素瘤的小鼠中的生物分布。 我们将调整我们建立的同基因转移模型， 细胞表达与人相同的分子定义的靶抗原 黑色素瘤细胞系M24 met和C8161形成了我们的黑色素瘤的基础， 在SCID小鼠中的转移模型。因此，B16黑色素瘤细胞将被稳定地 用编码人EGF-r的cDNA转染。我们的功效 降低肿瘤细胞生长速率的抗体-细胞因子融合蛋白 在我们的同基因黑色素瘤转移模型中， 与单克隆抗体、细胞因子或它们的混合物所达到的效果相比， 两者一旦单克隆抗体-细胞因子融合蛋白被证明是最有效的， 在这方面，参与这一进程的假定机制将是 考察这些将包括通过mAb-IL-2裂变的T细胞活化 通过mAb-TNF融合蛋白和肿瘤血管的破坏 proteins.一旦融合蛋白的功效及其作用机制被证实， 在完全免疫活性的鼠黑色素瘤中已经确立了作用， 转移模型，将努力建立这些参数， 我们在SCID小鼠中的人黑素瘤转移模型。这将涉及 用选定和确定的人类群体进行选择性重建 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 用于治疗转移性肿瘤的新方法的开发 黑素瘤

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.