Closing the system: production of viral antigen-presenting dendritic cells eliciting specific CD8+ T cell activation in fluorinated ethylene propylene cell culture bags

Journal of Translational Medicine

David Juncker¹  Veronique Laforte¹  Natalie Fekete²  Ariane V. Beland²  Corinne A. Hoesli³  Marie-Paule Lachambre⁴  Jean-Philippe Bastien⁴  Denis-Claude Roy⁵  Vibhuti Dave⁶ 

[1] Department of Biomedical Engineering, McGill University, Montreal, Québec, Canada;McGill Genome Centre, McGill University, Montreal, Québec, Canada;Department of Neurology and Neurosurgery, McGill University, Montreal, Québec, Canada;Department of Chemical Engineering, McGill University, Montreal, Québec, Canada;Department of Chemical Engineering, McGill University, Montreal, Québec, Canada;Department of Biomedical Engineering, McGill University, Montreal, Québec, Canada;Hematology-Oncology and Cell Therapy Institute, Hopital Maisonneuve-Rosemont Research Center, Montreal, Québec, Canada;Hematology-Oncology and Cell Therapy Institute, Hopital Maisonneuve-Rosemont Research Center, Montreal, Québec, Canada;Department of Medicine, Université de Montréal, Montreal, Québec, Canada;Hematology-Oncology and Cell Therapy Institute, Hopital Maisonneuve-Rosemont Research Center, Montreal, Québec, Canada;Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, Québec, Canada;

关键词: Cellular therapy;    Dendritic cell;    Fluorinated polymers;    Immunotherapy;    Monocyte;    Polystyrene;    Scale-down;   

DOI  :  10.1186/s12967-020-02543-1

来源: Springer

【 摘 要 】

BackgroundA major obstacle to anti-viral and -tumor cell vaccination and T cell immunotherapy is the ability to produce dendritic cells (DCs) in a suitable clinical setting. It is imperative to develop closed cell culture systems to accelerate the translation of promising DC-based cell therapy products to the clinic. The objective of this study was to investigate whether viral antigen-loaded monocyte-derived DCs (Mo-DCs) capable of eliciting specific T cell activation can be manufactured in fluorinated ethylene propylene (FEP) bags.MethodsMo-DCs were generated through a protocol applying cytokine cocktails combined with lipopolysaccharide or with a CMV viral peptide antigen in conventional tissue culture polystyrene (TCPS) or FEP culture vessels. Research-scale (< 10 mL) FEP bags were implemented to increase R&D throughput. DC surface marker profiles, cytokine production, and ability to activate antigen-specific cytotoxic T cells were characterized.ResultsMonocyte differentiation into Mo-DCs led to the loss of CD14 expression with concomitant upregulation of CD80, CD83 and CD86. Significantly increased levels of IL-10 and IL-12 were observed after maturation on day 9. Antigen-pulsed Mo-DCs activated antigen-responsive CD8+ cytotoxic T cells. No significant differences in surface marker expression or tetramer-specific T cell activating potency of Mo-DCs were observed between TCPS and FEP culture vessels.ConclusionsOur findings demonstrate that viral antigen-loaded Mo-DCs produced in downscaled FEP bags can elicit specific T cell responses. In view of the dire clinical need for closed system DC manufacturing, FEP bags represent an attractive option to accelerate the translation of promising emerging DC-based immunotherapies.

【 授权许可】

CC BY