CLINICAL SCALE EXPANSION OF HUMAN DENDRITIC CELLS

最近的临床研究表明，树突状细胞是一种强大的抗原提呈细胞，可在人类癌症患者体内诱导针对肿瘤抗原的免疫反应。然而，对这一新兴治疗方式的广泛评估和应用将需要可靠和有效的手段来产生足够数量的抗原负载树突状细胞，以便有效地对患者进行治疗。第一阶段计划定义了一种新的单程无血清介质灌流工艺，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树突状细胞。这一过程已在封闭的、人工操作的临床规模生物反应器系统中成功实施。通过在培养过程中测量乳酸随时间的变化，在临床规模上证明了非侵入性监测灌流过程的可行性。拟议的Il期研究的主要目标是优化细胞因子组合、介质灌注率、生物反应器生长面的材料组成和组织培养处理以及氧气输送，以在临床规模系统中最大限度地生产DC。在实现过程自动化以增强可靠性和重复性之后，在杜克大学医学中心进行的IDE批准的临床试验中，将对AstromReplicell(TM)系统中产生的DC进行安全性、生物分布和针对CEA-CAP-1肿瘤多肽抗原的免疫反应的诱导进行评估。拟议的商业应用：一个封闭的、自动化的、符合GMP的生物反应系统，用于树突状细胞的衍生、成熟、抗原负载和收获，将对癌症和传染病的免疫治疗具有重要价值。现有的获得树突状细胞的方法涉及多个开放的过程茎，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成功的基于树突状细胞的免疫疗法以可靠和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.