Abstract
BackgroundGenetic engineering of T-cells to express specific T cell receptors (TCR) has emerged as a novel strategy to treat various malignancies. More widespread utilization of these types of therapies has been somewhat constrained by the lack of closed culture processes capable of expanding sufficient numbers of T-cells for clinical application. Here, we evaluate a process for robust clinical grade manufacturing of TCR gene engineered T-cells.MethodsTCRs that target human papillomavirus E6 and E7 were independently tested. A 21 day process was divided into a transduction phase (7 days) and a rapid expansion phase (14 days). This process was evaluated using two healthy donor samples and four samples obtained from patients with epithelial cancers.ResultsThe process resulted in ~ 2000-fold increase in viable nucleated cells and high transduction efficiencies (64–92%). At the end of culture, functional assays demonstrated that these cells were potent and specific in their ability to kill tumor cells bearing target and secrete large quantities of interferon and tumor necrosis factor. Both phases of culture were contained within closed or semi-closed modules, which include automated density gradient separation and cell culture bags for the first phase and closed GREX culture devices and wash/concentrate systems for the second phase.ConclusionLarge-scale manufacturing using modular systems and semi-automated devices resulted in highly functional clinical-grade TCR transduced T-cells. This process is now in use in actively accruing clinical trials and the NIH Clinical Center and can be utilized at other cell therapy manufacturing sites that wish to scale-up and optimize their processing using closed systems.
Highlights
Genetic engineering of T-cells to express specific T cell receptors (TCR) has emerged as a novel strategy to treat various malignancies
Mononuclear apheresis was obtained from healthy donors or patients with human papillomavirus (HPV)-16+ epithelial cancer
A consistent pattern was observed among the six samples tested regardless of whether samples were from healthy donors or patients, or when differing numbers of cells were used at cryopreservation
Summary
Genetic engineering of T-cells to express specific T cell receptors (TCR) has emerged as a novel strategy to treat various malignancies. Much of the recent attention that has led to this increased development has been due to encouraging phase I/II clinical trials using CAR T-cells directed against CD19 and CD22 [1,2,3] and TCR-modified T-cells directed against NY-ESO-1, MAGE, and GP100 [4,5,6,7]. Both of these approaches have their own particular nuances. While TCR-transduced T-cells are able to recognize intracellular antigens processed by major histocompatibility (MHC) proteins, CAR T-cells only target surface antigens, but these can be recognized in a MHC-independent fashion, allowing more patients to be treated with a specific viral vector.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
