EAGER: Biomanufacturing: Polymer Protective Effector T-Cell Isolation and Centrifugal Bioreactor Expansion for a Parasitic Disease Model with Relevance in Human Cancer Treatment

最近的成功表明免疫系统可以用来对抗癌症。细胞毒性免疫细胞能够对抗多种癌症的发现，为对抗癌症提供了新的途径。在开发利用这些知识来改进治疗癌症的方法方面，仍然存在两个挑战。第一个是开发治疗方法，阻止癌细胞干扰细胞毒性免疫细胞的杀伤能力。第二种方法是分离和扩增免疫细胞，以便及时用于治疗危及生命的癌症患者。这些细胞在血液和癌症组织中出现的频率很低。化学工程和兽医免疫学的研究人员已经联合起来应对这些挑战。细胞毒性细胞将被保护性塑料涂层隔离，而其他细胞则被破坏。一种离心式生物反应器已被开发用于探索快速扩增培养免疫细胞的方法。对一种在牛中引起疾病的蜱虫寄生虫的研究已经确定了一个模型系统，该系统提供了一个持续的细胞毒性免疫细胞来源，以对抗行为类似白血病细胞的寄生虫感染的靶细胞。在优化快速扩增细胞的方法的同时，可以探索分离细胞毒性免疫细胞的方法，这将是分离癌症特异性免疫细胞所需要的。将测试优化培养条件的方法。扩增培养的免疫细胞可以监测其纯度和细胞毒性活性。这项研究的结果将很容易适用于来自癌症患者的细胞毒性免疫细胞。t淋巴细胞免疫疗法提供了新的方法来对抗入侵和接管细胞机制，导致恶性肿瘤的癌症。该项目强调使用一种新的平台来评估分离和快速扩增细胞毒性T细胞的方法。该模型使用了一种由蜱虫传播的原生动物寄生虫，即小孢子虫或小孢子虫。在摄取血粉的过程中，感染形式侵入淋巴细胞。在进入淋巴细胞后，寄生虫劫持细胞分裂的调节机制，导致细胞与寄生虫同步分裂。受感染细胞产生的细胞因子失调会干扰免疫系统在受感染细胞快速增殖之前产生保护性反应的能力，导致受感染动物除非得到治疗否则死亡。最近的研究表明，针对疟原虫的细胞毒性T细胞可用于体外杀死受感染的细胞。癌症免疫治疗需要的是分离特异性效应t细胞的方法，以靶向被感染的快速分裂的恶性细胞，并有效地扩增这些细胞，使其产生大量的细胞，以输注回患者体内，以提高生存率。在这个项目中，将结合几种技术来创造大量的CD8 T淋巴细胞（CTL）来对抗细小T虫：1)通过用细小T抗原脉冲树突状细胞（主要的抗原呈递细胞）刺激外周血T细胞来激活接种牛的抗疾病CTL； 2)使用一种新的聚合物保护涂层技术从培养物中选择性地分离出特定的CTL，所有不需要的细胞都通过裂解去除；3)检测ctl的克隆和维持细胞毒性活性；4)利用新型离心式生物反应器（CBR）大量扩增CTL。将获得与CTL激活有关的新知识，这些知识对对抗多种癌症有用。聚合物保护涂层技术将从目前的重点用于干细胞分离扩展到CTL。CTL细胞毒性试验将被改进，特别是使用T. parva模型。将首次获得扩展CBR以扩展CTL用于免疫治疗的认识。

1645249 - Van WieRecent successes have shown the immune system can be used to fight cancers. The finding that cytotoxic immune cells develop against many types of cancer is offering fresh ways to fight cancer. Two challenges remain in developing ways to exploit this knowledge to improve methods for treating cancers. The first is developing therapies to block the ability of the cancer cells to interfere with killing by the cytotoxic immune cells. The second is isolating and expanding the immune cells for timely use to treat patients with life threatening cancers. The frequency of these cells is low in blood and cancer tissue. Investigators in chemical engineering and veterinary immunology have joined together to address these challenges. Cytotoxic cells will be separated by protective plastic coatings while the other cells are disrupted. A centrifugal bioreactor has been developed to explore methods for rapidly expanding cultures of immune cells. Study of a tick born parasite causing disease in cattle has identified a model system that provides a constant source of cytotoxic immune cells against parasite infected target cells that behave like leukemia cells. Methods for isolating the cytotoxic immune cells can be explored while optimizing methods to rapidly expand the cells, as will be needed for isolating cancer specific immune cells. Methods for optimizing culture conditions will be tested. The expanded cultures of immune cells can be monitored for purity and cytotoxic activity. Results from the research will be readily applicable for use with cytotoxic immune cells from cancer patients.T-lymphocyte immunotherapy is offering fresh ways to fight cancers that invade and take over cell machinery, leading to malignant tumors. This project emphasizes the use of a novel platform for evaluating methods to isolate and rapidly expand cytotoxic T cells. The model involves use of a protozoan parasite, Theileria parva or T. parva, transmitted by ticks. The infectious form invades lymphocytes following introduction during uptake of a blood meal. Following entrance into a lymphocyte, the parasite hijacks the regulatory mechanism for cell division causing cells to divide in synchrony with the parasite. Dysregulation of cytokine production by the infected cells interferes with the capacity of the immune system's capacity to develop a protective response before rapid proliferation of the infected cells leading to death of infected animals unless treated. Recent research shows cytotoxic T cells programmed against the T. parva parasite can be used to kill infected cells ex vivo. What is needed for cancer immunotherapy are ways to isolate the specific effector T-cells for targeting infected rapidly-dividing malignant cells, and to efficiently expand these cells to manufacture very large numbers for infusion back into a patient to increase survival. Several technologies will be combined in this project to create large numbers of effector CD8 T-lymphocytes (CTL) against T. parva to: 1) activate disease fighting CTL from vaccinated cattle, by stimulating peripheral blood T cells with T parva antigen-pulsed dendritic cells (the major antigen presenting cells; 2) selectively isolate specific CTL from the culture using a new polymer protective coating technique where all unwanted cells are removed by lysis; 3) test CTLs for clonality and maintenance of cytotoxic activity; and 4) expand CTL to large numbers using a novel centrifugal bioreactor (CBR). New knowledge will be gained relevant to activation of CTL useful in combating a variety of cancers. The polymer protective coating technique will be expanded from its current emphasis for use in isolating stem cells to CTL. CTL cytotoxicity assays will be refined especially by using the T. parva model. Understanding will be gained on extending the CBR for the first time to expand CTL for immunotherapy.