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

Washington State University, Pullman WA

Investigators

Abstract

1645249 - Van Wie Recent 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.