GPI-ANCHORED MOLECULES AS THERAPEUTIC VACCINES

描述(改编自申请人的摘要)：经以下方法修饰的肿瘤 对B7.1等共刺激分子的基因转染现在正在进行中 被认为是一种潜在的治疗性肿瘤疫苗。然而，转染法 并不总是有效的并且对于许多细胞类型来说可能是困难的， 尤其是从患者身上新分离的肿瘤细胞。调查员 建议确定共刺激基因修饰的肿瘤的疗效 分子使用一种新的方法。研究表明，纯化的糖基 磷脂酰肌醇(GPI)锚定的细胞表面蛋白可以 通过与蛋白质孵育自发地结合到细胞中 细胞。这种独特的性质可以用来重建细胞表面 在细胞膜上表达受体而不需要使用基因转染法。 利用重组技术，他们构建并表达了 纯化的GPI-B7.1分子锚定在肿瘤上。调查员提议 确定重组的肿瘤的免疫治疗潜力 GPI-B7诱导肿瘤特异性免疫。利用体外人类 实验和体内小鼠实验，诱导的最佳条件 肿瘤免疫力将被确定。 他们还将确定细胞因子是否可以增强诱导的免疫 通过GPI锚定分子修饰的肿瘤。人类肿瘤细胞系将 被开发来确定GPI-B7.1重组的疗效 自体肿瘤特异性CTL的体外培养。其他共刺激作用 GPI-ICAM-1和热稳定抗原等黏附分子将 单独开发和测试，或与B7.1结合使用，以诱导 肿瘤特异性免疫。这项拟议研究的结果将 展示GPI锚定的有效性和潜在应用 分子修饰肿瘤作为肿瘤免疫治疗的治疗性疫苗。 从长远来看，从这项研究中获得的知识可以用来 研发一种治疗癌症的“人造细胞疫苗”。

DESCRIPTION (Adapted from the Applicant's Abstract): Tumors modified by transfecting genes for co-stimulatory molecules such as B7.1 are now being considered as a potential therapeutic tumor vaccine. However, transfection is not always efficient and can be difficult with many cell types, especially freshly isolated tumor cells from patients. The investigator proposes to determine efficacy of tumors modified with co-stimulatory molecules using a novel approach. Studies have shown that purified glycosyl phosphatidyl inositol (GPI)-anchored cell surface proteins can be spontaneously incorporated into cells by incubating the proteins with the cells. This unique property can be used to reconstitute cell surface expression receptors on cell membranes without the use of gene transfection. Using recombinant techniques, they have constructed and expressed the purified GPI-anchored B7.1 molecule on tumors. The investigator proposes to determine the immunotherapeutic potential of tumors reconstituted with GPI-B7 in inducing tumor specific immunity. Using in vitro human experiments and in vivo mouse experiments, optimal conditions for inducing tumor immunity will be determined. They will also determine whether cytokines can enhance the immunity induced by tumors modified with GPI-anchored molecules. Human tumor cell lines will be developed to determine the efficacy of GPI-B7.1 reconstitution on autologous tumor specific CTL development in vitro. Other co-stimulatory adhesion molecules such as GPI-ICAM-1 and heat-stable antigen will be developed and tested alone or in combination with B7.1, for the induction of tumor specific immunity. The results from the proposed research will demonstrate the efficacy and potential application of the GPI-anchored molecule modified tumors as a therapeutic vaccine for tumor immunotherapy. In the long term the knowledge obtained from this study could be used to develop an "artificial cell vaccine" to treat cancer.