Melanoma antigens recognized by B and T helper lymphocytes

这项提案的主要目标是确定黑色素瘤相关抗原 这些疫苗是这些患者主动免疫治疗的候选疫苗。 抗原将由患者的B细胞确定。 具体地说， 免疫原性抗原将通过组合抗体（Fab） 来自患者淋巴细胞并在表面表达的文库 丝状真菌的 这种方法有许多潜在的优点 相对于使用来自于以下的单克隆抗体（mAb）的常规方法， 杂交瘤或Epstein巴尔病毒（EBV）转化的B细胞。 优惠 通过黑素瘤细胞表达组合Fab选择的结构， 与正常组织相比，这表明它们可用作 病人对肿瘤的体液免疫 鉴于此前 辅助性和细胞溶解性T细胞（CTL）表位的演示 除了最初由B细胞定义的抗原外，选择的抗原也可以诱导 患者的细胞免疫。 具体目标是：1）表达 来源于黑色素瘤患者B细胞的组合Fab B文库， 使用pComb-3 H载体将丝状真菌（M13）的表面进行修饰，并选择 那些与黑色素瘤优先表达的蛋白抗原结合的Fab 细胞与正常细胞相比; 2）表征、克隆和表达 由选择的组合Fab定义的蛋白抗原;和3）确定 所选抗原与患者T细胞的反应性。 选定的抗原可用于未来的研究，用于主动免疫治疗。 黑色素瘤患者。

The major goal of this proposal is to identify melanoma-associated antigens that are candidate vaccines for active immunotherapy of these patients. The antigens will be defined by patients' B cells. Specifically, immunogenic antigens will be identified by combinatorial antibody (Fab) libraries derived from patients' lymphocytes and expressed on the surface of filamentous phages. This approach has numerous potential advantages over conventional approaches using monoclonal antibodies (mAb) derived from hybridomas or Epstein Barr Virus (EBV)-transformed B cells. Preferential expression of combinatorial Fab-selected structures by melanoma cells as compared to normal tissues would suggest their usefulness as modulators of patients' humoral immunity to their tumors. In light of previous demonstrations of both helper and cytolytic T-cell (CTL) epitopes on antigens originally defined by B cells, selected antigens also may induce cellular immunity in patients. The specific aims are to; 1) express combinatorial Fab libraries derived from melanoma patients' B cells on the surface of filamentous phages (M13) using the pComb-3H vector, and select those Fab binding to protein antigens preferentially expressed by melanoma cells as compared to normal cells; 2) characterize, clone and express protein antigens defined by selected combinatorial Fab; and 3) determine the reactivities of selected antigens with patients' T cells. Selected antigens may be used in future studies for active immunotherapy of melanoma patients.