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

1645249-Van Wie最近的成功表明免疫系统可以用来抗击癌症。细胞毒性免疫细胞可以抵抗多种癌症，这一发现为抗击癌症提供了新的方法。在开发利用这一知识来改进癌症治疗方法的方法方面仍然存在两个挑战。首先是开发治疗方法，以阻断癌细胞干扰细胞毒性免疫细胞杀戮的能力。第二种方法是分离和扩大免疫细胞，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的频率很低。化学工程和兽医免疫学的研究人员已经联手应对这些挑战。细胞毒性细胞将被保护性塑料涂层隔开，而其他细胞则被破坏。开发了一种离心式生物反应器，以探索快速扩增免疫细胞培养的方法。对一种扁虱传播的寄生虫在牛身上致病的研究已经确定了一种模型系统，该系统可以提供持续的细胞毒性免疫细胞来源，以对抗寄生虫感染的靶细胞，这些细胞的行为类似于白血病细胞。可以探索分离细胞毒性免疫细胞的方法，同时优化快速扩增细胞的方法，这将是分离癌症特异性免疫细胞所必需的。将测试优化培养条件的方法。免疫细胞的扩增培养可以监测其纯度和细胞毒活性。这项研究的结果将很容易应用于癌症患者的细胞毒性免疫细胞。T淋巴细胞免疫疗法正在提供新的方法来对抗入侵并接管细胞机器的癌症，从而导致恶性肿瘤。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。该模型使用了一种原生动物寄生虫--微小泰勒虫或微小锥虫，这种寄生虫是通过扁虱传播的。在进食血餐时，这种感染形式会侵入淋巴细胞。在进入淋巴细胞后，寄生虫劫持了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产生的失调干扰了免疫系统在感染细胞快速增殖导致受感染动物死亡之前产生保护性反应的能力，除非得到治疗。最近的研究表明，针对弓形虫寄生虫编程的细胞毒性T细胞可以用来在体外杀死感染的细胞。癌症免疫治疗需要的是分离特定的效应T细胞，以靶向受感染的快速分裂的恶性细胞，并有效地扩增这些细胞，制造大量的细胞，然后回输给患者，以提高存活率。该项目将结合多种技术创建大量抗微小毛滴虫的效应者CD8 T淋巴细胞(CTL)：1)通过用T parva抗原致敏的树突状细胞(主要的抗原提呈细胞)刺激外周血T细胞，激活免疫牛的抗病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特定的CTL，通过裂解去除所有不需要的细胞；3)测试CTL的克隆性和维持细胞毒活性；以及4)使用新型的离心式生物反应器(CBR)大量扩增CTL。将获得与激活CTL相关的新知识，这些CTL有助于对抗各种癌症。聚合物保护性涂层技术将从目前用于分离干细胞的重点扩展到CTL。CTL细胞毒性测定将得到改进，特别是通过使用T.parva模型。双方将就首次扩大CBR以扩大CTL用于免疫治疗达成谅解。

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.