CLINICAL SCALE EXPANSION OF HUMAN DENDRITIC CELLS

最近的临床研究表明，树突状细胞是有效的抗原呈递细胞，可诱导人类癌症患者针对肿瘤抗原的免疫反应。然而，对这种新兴治疗方式的广泛评估和应用将需要一种可靠且有效的方法来产生足够数量的负载抗原的树突状细胞，以进行有效的患者治疗。第一阶段计划定义了一种新颖的单次无血清培养基灌注工艺，用于从患者的白细胞分离细胞中以高接种密度生产高功能的树突状细胞。该过程已在封闭的手动操作的临床规模生物反应器系统中成功实施。通过在培养过程中随时间测量乳酸来非侵入性监测灌注过程的可行性已在临床规模得到证实。所提出的II期研究的主要目标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输送，以在临床规模系统中最大程度地产生DC。在实现流程自动化以提高可靠性和可重复性后，将在杜克大学医学中心进行 IDE 批准的临床试验，评估 AastromReplicell(TM) 系统中生成的 DC 的安全性、生物分布和诱导针对 CEA-CAP-1 肿瘤肽抗原的免疫反应。拟议的商业应用：用于树突状细胞衍生、成熟、抗原加载和收获的封闭、自动化、符合 GMP 的生物反应器系统对于癌症和传染病的免疫治疗具有巨大价值。现有的树突状细胞衍生方法涉及多个开放过程干，以及相关的设备和劳动力成本。全自动 DC 培养过程的开发将使基于树突状细胞的免疫疗法以可靠且具有成本效益的方式得到广泛应用。

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.