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An accelerated, clinical-grade protocol to generate high yields of type 1-polarizing messenger RNA-loaded dendritic cells for cancer vaccination.
- Source :
-
Cytotherapy [Cytotherapy] 2018 Sep; Vol. 20 (9), pp. 1164-1181. Date of Electronic Publication: 2018 Aug 16. - Publication Year :
- 2018
-
Abstract
- Background: Many efforts have been devoted to improve the performance of dendritic cell (DC)-based cancer vaccines. Ideally, a DC vaccine should induce robust type 1-polarized T-cell responses and efficiently expand antigen (Ag)-specific cytotoxic T-cells, while being applicable regardless of patient human leukocyte antigen (HLA) type. Production time should be short, while maximally being good manufacturing practice (GMP)-compliant. We developed a method that caters to all of these demands and demonstrated the superiority of the resulting product compared with DCs generated using a well-established "classical" protocol.<br />Methods: Immunomagnetically purified monocytes were cultured in a closed system for 3 days in GMP-compliant serum-free medium and cytokines, and matured for 24 h using monophosphoryl lipid A (MPLA)+ interferon-gamma (IFN-γ). Mature DCs were electroporated with messenger RNA (mRNA) encoding full-length antigen and cryopreserved. "Classical" DCs were cultured for 8 days in flasks, with one round of medium and cytokine supplementation, and matured with tumor necrosis factor alpha (TNF-α) + prostaglandin E2 (PGE2) during the last 2 days.<br />Results: Four-day MPLA/IFN-γ-matured DCs were superior to 8-day TNF-α/PGE2-matured DCs in terms of yield, co-stimulatory/co-inhibitory molecule expression, resilience to electroporation and cryopreservation and type 1-polarizing cytokine and chemokine release after cell thawing. Electroporated and cryopreserved DCs according to our protocol efficiently present epitopes from tumor antigen-encoding mRNA, inducing a strong expansion of antigen-specific CD8+ T-cells with full cytolytic capacity.<br />Conclusion: We demonstrate using a GMP-compliant culture protocol the feasibility of generating high yields of mature DCs in a short time, with a superior immunogenic profile compared with 8-day TNF-α/PGE2-matured DCs, and capable of inducing vigorous cytotoxic T-cell responses to antigen from electroporated mRNA. This method is now being applied in our clinical trial program.<br /> (Copyright © 2018 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)
- Subjects :
- Antigens, Neoplasm genetics
Antigens, Neoplasm immunology
Cell Differentiation
Cryopreservation
Dendritic Cells immunology
Dinoprostone pharmacology
Electroporation
Epitopes
Humans
Interferon-gamma pharmacology
Lipid A analogs & derivatives
Lipid A pharmacology
Monocytes cytology
T-Lymphocytes, Cytotoxic immunology
Tumor Necrosis Factor-alpha pharmacology
Cancer Vaccines
Cell Culture Techniques methods
Dendritic Cells cytology
RNA, Messenger genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1477-2566
- Volume :
- 20
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Cytotherapy
- Publication Type :
- Academic Journal
- Accession number :
- 30122654
- Full Text :
- https://doi.org/10.1016/j.jcyt.2018.06.006