CLINICAL SCALE EXPANSION OF HUMAN DENDRITIC CELLS

最近的临床研究表明，树突状细胞是一种有效的抗原呈递细胞，可诱导人类癌症患者对肿瘤抗原的免疫应答。然而，这种新兴治疗方式的广泛评价和应用将需要可靠且有效的手段来产生足够量的抗原负载的树突状细胞以用于有效的患者治疗。I期项目定义了一种新的单程无血清培养基灌注工艺，用于以高接种密度从患者的白细胞去除术细胞中生产高功能性树突状细胞。该工艺已在封闭、手动操作的临床规模生物反应器系统中成功实施。在培养过程中，通过乳酸盐测量随时间的推移对灌注过程进行无创监测的可行性已在临床规模上得到证实。所提出的II期研究的主要目的是优化细胞因子组合、培养基灌注速率、材料组成和生物反应器生长表面的组织培养处理以及氧气递送，以在临床规模系统中最大限度地产生DC。在提高可靠性和再现性的过程自动化之后，在杜克大学医学中心的IDE批准的临床试验中，将评估AastromReplicellTM系统中产生的DC的安全性、生物分布和对CEA-CAP-1肿瘤肽抗原的免疫应答的诱导。拟定商业应用：一个封闭的、自动化的、符合GMP的生物反应器系统用于树突状细胞的衍生、成熟、抗原加载和收获，对于癌症和感染性疾病的免疫治疗具有重要价值。现有的树突状细胞衍生方法涉及多个开放式工艺系统，具有相关的设备和劳动力成本。全自动DC培养过程的开发将使成功的基于树突状细胞的免疫疗法以可靠和成本有效的方式广泛应用。

Recent clinical studies have demonstrated that dendritic cells are potent antigen presenting cells for induction of immune responses against tumor antigens in human cancer patients. However, widespread evaluation and application of this emerging treatment modality will require a reliable and efficient means to generate sufficient quantities of antigen-loaded dendritic cells for effective patient therapy. The Phase I program has defined a novel single-pass serum-free medium perfusion process for production of highly functional dendritic cells at high inoculum density from patients' leukapheresis cells. This process has been implemented successfully in a closed, manually operated, clinical scale bioreactor system. Feasibility for non-invasive monitoring of the perfusion process by lactate measurement with time during culture has been demonstrated at clinical scale. The primary objectives of the proposed Phase Il studies are to optimize cytokine combinations, medium perfusion rates, material composition and tissue culture treatment of the bioreactor growth surface and oxygen delivery for maximum production of DCs in the clinical scale system. After automation of the process to enhance reliability and reproducibility, DCs generated in the AastromReplicell(TM) System will be evaluated for safety, biodistribution and induction of immune responses against CEA- CAP-1 tumor-peptide antigen under IDE-approved clinical trials at Duke University Medical Center. PROPOSED COMMERCIAL APPLICATION: A closed, automated, GMP-compliant bioreactor system for dendritic cell derivation, maturation, antigen-loading and harvest would be of great value for immunotherapy of cancer and infectious diseases. The existing methods for derivation of dendritic cells involve multiple open process stems with associated costs in equipment and labor. The development of a fully automated DC culture process will enable widespread application of successful dendritic cell-based immunotherapies in a reliable and cost- effective fashion.