Isolation of novel tumor antigens

描述(由申请人提供)： 体内递送的质粒DNA表达载体已被证明是一种 诱导针对所表达基因的免疫反应的有效手段 序列。用这种方法刺激宿主对肿瘤的免疫反应 是一种很有吸引力的癌症治疗方法。基因治疗的有效性 免疫治疗将取决于肿瘤相关抗原的特性。 (TAA)用于诱导抗肿瘤免疫反应，以及部位和 TAA的表达水平。米卢斯？血管内非病毒基因传递 技术已经证明在诱导转基因导向的方面特别有效 免疫反应，取决于被转染者的细胞类型(包括 抗原提呈细胞)和获得的高水平表达。 这项拨款提案的主要目的是利用这些被证明是高效的 基因转移技术在相关体内肿瘤模型中评估TAAs， 从人肿瘤表达文库中筛选新的TAA。在第一阶段，我们 将确定这些体内非病毒基因转移技术是否会导致 B16小鼠的预防性和治疗性抗肿瘤免疫反应 黑色素瘤模型。同时，我们将确定最佳的基因疫苗接种。 条件(例如，细胞因子基因的联合传递、最低限度的pDNA 必填项)。在第二阶段，将构建消减表达文库 从人类肿瘤细胞中分离出来。非法抗肿瘤文库中的TAA基因 将通过传输单个克隆或(部分)来选择响应 将文库送入小鼠，随后进行肿瘤挑战。经过连续几轮， 将隔离特定的和高活动的TAA。成功的选拔和 由于非常有效的免疫，可以对新的TAA进行评估 在Mirus之后得到的回应是什么？血管内基因传递技术， 它也可以用来向癌症患者运送TAA。

DESCRIPTION (provided by applicant): In vivo delivery of plasmid DNA expression vectors has shown to be an efficient means of inducing an immune response against the expressed gene sequence. Stimulating host immune responses against tumors using this method is an attractive cancer therapy approach. The effectiveness of genetic immunotherapy will depend on the properties of the tumor-associated antigen (TAA) used to induce the anti-tumor immune response, as well as the site and level of TAA expression. Mirus? intravascular non-viral gene delivery techniques have shown especially efficient at inducing transgene-directed immune responses, due to the types of cells that are transfected (including antigen presenting cells) and the high levels of expression that are obtained. The main Aim of this grant proposal is to use these proven highly efficient gene transfer techniques to evaluate TAAs in relevant in vivo tumor models, and to select novel TAAs from human tumor expression libraries. In Phase I, we will determine if these in vivo non-viral gene transfer techniques result in prophylactic and therapeutic anti-tumor immune responses in the B16 murine melanoma model. In parallel, we will determine the optimal genetic vaccination conditions (e.g., co-delivery of cytokine genes, minimal amount of pDNA required). In Phase II, subtracted expression libraries will be constructed from human tumor cells. TAA genes in the library that illicit anti-tumor responses will be selected by transferring individual clones or (partial) libraries into mice, followed by tumor challenge. Through successive rounds, specific and highly active TAAs will be isolated. Successful selection and evaluation of novel TAAs can be performed because of the very efficient immune response obtained following Mirus? intravascular gene delivery techniques, which can also be used to deliver TAAs to cancer patients.