PILOT--MODIFICATION OF MELANOMA CELLS WITH GPI-ANCHORED IL-2

许多研究表明，细胞因子在细胞凋亡中起主要作用。 恶性肿瘤如黑色素瘤的发病机制和治疗。系统性 在肿瘤患者中施用某些细胞因子已经显示， 增强肿瘤特异性免疫应答和某些肿瘤的消退 患者用细胞因子特异性基因转染的肿瘤已经被发现， 显示诱导肿瘤特异性免疫，表明肿瘤细胞 表达细胞因子将具有潜在的治疗价值， 疫苗。我们建议测试一种方法的有效性， 作为将细胞因子分子导入肿瘤的替代方案， 其中基因转染是不可行或不希望的。该方法 使用纯化的分子，可以在几个小时内完成， 合适的试剂。根据这项建议，我们将具体： 1.糖基磷脂酰肌醇（GPI）的构建、表达和纯化。 锚定IL-2; 2.确定黑色素瘤细胞是否用纯化的GPI重建， IL-2可以诱导肿瘤特异性免疫。GPI锚定的细胞因子将是一种 替代现有的分子，因为它将不同于 母体分子在其自发结合到 疏水/两亲表面，如细胞膜。知识 从这个种子赠款提案中获得的将是巨大的价值， 制备用于治疗皮肤癌的免疫原性肿瘤细胞， 黑素瘤

Many studies have shown that cytokines play a major role in the pathogenesis and cure of malignant tumors such as melanoma. Systemic administration of certain cytokines in tumor patients have shown to augment tumor specific immune responses and regression of tumors in some patients. Tumors transfected with genes specific for cytokines have been shown to induce tumor specific immunity suggesting that tumor cells expressing cytokines will be of potential therapeutic value as tumor vaccines. We propose to test the efficacy of a method that could be used as an alternative for introduction of cytokine molecules onto tumors, where gene transfection is not feasible or desirable. The proposed method uses purified molecules and can be completed in a matter of hours with appropriate reagents. Under this proposal specifically we will: 1. construct, express and purify glycosyl-phosphatidylinositol (GPI)- anchored IL-2; 2. determine whether the melanoma cells reconstituted with purified GPI- IL-2 can induce tumor specific immunity. GPI-anchored cytokines will be an alternative to the existing molecules since it will be different from the parent molecule in terms of its ability to spontaneously incorporate into hydrophobic/amphiphilic surfaces such as cell membranes. The knowledge obtained from this seed grant proposal will be of immense value in preparing immunogenic tumor cells for treatment of skin cancers such as melanoma.