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  3. Closing The System: Production of Viral Antigen-Presenting Dendritic Cells Eliciting Specific CD8+ T Cell Activation in Fluorinated Ethylene Propylene Cell Culture Bags

Closing The System: Production of Viral Antigen-Presenting Dendritic Cells Eliciting Specific CD8+ T Cell Activation in Fluorinated Ethylene Propylene Cell Culture Bags

November 06th, 2020

Peer-Reviewed Publication from Journal of Translational Medicine

By: Jean‑Philippe Bastien, Natalie Fekete, Ariane V. Beland, Marie‑Paule Lachambre, Veronique Laforte, David Juncker, Vibhuti Dave, Denis‑Claude Roy and Corinne A. Hoesli

For full article click here

Abstract

Background: A 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 specifc T cell activa‑ tion can be manufactured in fuorinated ethylene propylene (FEP) bags.

Methods: Mo-DCs were generated through a protocol applying cytokine cocktails combined with lipopolysac‑ charide or with a CMV viral peptide antigen in conventional tissue culture polystyrene (TCPS) or FEP culture ves‑ sels. Research-scale (<10 mL) FEP bags were implemented to increase R&D throughput. DC surface marker profles, cytokine production, and ability to activate antigen-specifc cytotoxic T cells were characterized.

Results: Monocyte diferentiation into Mo-DCs led to the loss of CD14 expression with concomitant upregulation of CD80, CD83 and CD86. Signifcantly 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 signifcant diferences in surface marker expression or tetramer-specifc T cell activating potency of Mo-DCs were observed between TCPS and FEP culture vessels.

Conclusions: Our fndings demonstrate that viral antigen-loaded Mo-DCs produced in downscaled FEP bags can elicit specifc T cell responses. In view of the dire clinical need for closed system DC manufacturing, FEP bags repre‑ sent an attractive option to accelerate the translation of promising emerging DC-based immunotherapies.

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