Melanoma antigens recognized by B and T helper lymphocytes

这项提议的主要目标是识别黑色素瘤相关抗原。 这些疫苗是这些患者主动免疫治疗的候选疫苗。 这些抗原将由患者的B细胞来定义。具体来说， 免疫原性抗原将通过组合抗体(Fab)进行鉴定 来源于患者淋巴细胞并在表面表达的文库 丝状噬菌体。这种方法有许多潜在的优势。 与传统方法相比，使用来源于 杂交瘤或EB病毒(EBV)转化的B细胞。优惠 黑色素瘤细胞表达Fab选择的组合结构 与正常组织相比可能表明它们作为调节因子的作用 患者对肿瘤的体液免疫。根据以前的情况 辅助性和溶细胞性T细胞(CTL)表位的研究 最初由B细胞定义的抗原，选定的抗原也可能诱导 患者的细胞免疫功能。具体目的是：1)表达 来自黑色素瘤患者B细胞的组合Fab文库 利用pComb-3H载体获得丝状噬菌体(M13)的表面，并选择 与黑色素瘤优先表达的蛋白抗原结合的Fab 与正常细胞相比；2)鉴定、克隆和表达 由选定的组合Fab定义的蛋白质抗原；以及3)确定 选定的抗原与患者T细胞的反应性。 选定的抗原可用于未来的主动免疫治疗的研究 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.