EXPANSION AND GENETIC TRANSDUCTION OF EBV-SPECIFIC CTLS

EBV 特异性 CTLS 的扩增和遗传转导

• 批准号: 2867484
• 负责人: DOUGLAS Matthew SMITH
• 金额: $ 9.98万
• 依托单位: AASTROM BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-15 至 2000-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2867484
• 关键词: Epstein Barr virus; Hodgkin's disease; biomedical equipment development; bioreactors; cytotoxic T lymphocyte; dendritic cells; genetic markers; genetic transduction; lymphoblast; mass tissue /cell culture; transfection /expression vector; virus antigen

Recent studies have demonstrated the EBV-specific CTLs are highly effective for prophylaxis of therapy of EBV-lymphoma in immunocompromised patients. However, widespread application of this emerging treatment strategy and extension to other EBV- associated lymphoproliferative diseases (LPDs) will require a reliable and efficient means to generate sufficient quantities of tumor antigen-specific CTLs for effective patient therapy. The primary objective of the proposed studies is to develop and implement a unique process for ex vivo derivation, antigen- specific expansion in response to autologous EBV-transformed lymphoblastoid cell lines or LMP2a-transduced dendritic cells, and retroviral-mediated gene-marking of EBV-specific CTLs in a perfused clinical-scale bioreactor system. The hypothesis that continuous single-pass medium exchange will enable enhanced T- cell production, function, and genetic transduction using flow- through technology in the AastromReplicell TM Cell Production System will be tested. The Phase II program will focus on optimization and full automation of the processes in compliance with GMP regulations. The automated bioreactor system will be tested in clinical trials to assess the safety and efficacy of gene-marked EBV-specific CTLs for therapy of EBV-associated lymphoma and Hodgkin s disease. An automated clinical scale bioreactor system would enable efficient and reliable production of highly functional gene-marked T-cells for antigen-specific immunotherapy of EBV-associated LPDs and other cancers. PROPOSED COMMERCIAL APPLICATION A closed, GMP-compliant, bioreactor system for ex vivo derivation, expansion and gene-marking of therapeutic quantities of EBV-specific CTLs would have immediate commercial value for prevention and treatment of EBV-associated LPDs in immunocompromised patients. The development of an automated process for antigen-specific T-cell expansion will reduce open- process steps and associated costs in equipment and labor, thus enabling widespread application of T-cell-based cancer immunotherapies in a reliable and cost-effective fashion.

最近的研究表明，EBV特异性CTL对免疫低下患者的EBV淋巴瘤的预防治疗是非常有效的。然而，这一新兴治疗策略的广泛应用以及推广到其他EBV相关淋巴增殖性疾病(LPD)将需要可靠和有效的手段来产生足够数量的肿瘤抗原特异性CTL，以便有效地进行患者治疗。这项研究的主要目的是开发和实施一种独特的方法，用于体外衍生、抗原特异性扩增以响应自体EBV转化的淋巴母细胞系或LMP2a转导的树突状细胞，以及在灌流的临床规模的生物反应器系统中逆转录病毒介导的EBV特异性CTL的基因标记。持续的单次介质交换将在AstromReplicell TM细胞生产系统中使用流通式技术增强T细胞的生产、功能和遗传转导的假设将得到验证。第二阶段计划将重点放在符合GMP规定的过程的优化和完全自动化上。该自动生物反应器系统将在临床试验中进行测试，以评估基因标记的EB病毒特异性CTL治疗EB病毒相关淋巴瘤和霍奇金S病的安全性和有效性。自动化的临床规模生物反应器系统将能够高效和可靠地生产高功能的基因标记T细胞，用于EBV相关LPD和其他癌症的抗原特异性免疫治疗。建议的商业应用一个封闭的、符合GMP的生物反应器系统，用于体外衍生、扩增和基因标记治疗量的EBV特异性CTL，将对预防和治疗免疫低下患者的EBV相关LPD具有直接的商业价值。抗原特异性T细胞扩增自动化过程的开发将减少开放过程步骤以及设备和劳动力的相关成本，从而使以T细胞为基础的癌症免疫疗法能够以可靠和具有成本效益的方式广泛应用。