An accelerated, clinical-grade protocol to generate high yields of type 1-polarizing messenger RNA–loaded dendritic cells for cancer vaccination

Brabants, Elisabeth; Heyns, Kelly; Smet, Steven; Devreker, P.; Ingels, Joline; Cabooter, Nancy De; Debacker, Veronique; Dullaers, Mélissa; Meerbeeck, Jan P. van; Vandekerckhove, Bart; Vermaelen, Karim

doi:10.1016/j.jcyt.2018.06.006

Cited by 13 publications

(14 citation statements)

References 52 publications

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“…Finally, the use of MPLA instead of LPS in the maturation cocktail needed to be optimized. It has already been shown that the use of MPLA/IFNγ to mature moDC was efficient to induce functional cells [14][15][16] and results presented here confirmed this observation. The use of MPLA/IFNγ led to higher secretion of IL12p70 by mature moDC than with LPS/IFNγ.…”

Section: Discussionsupporting

confidence: 88%

“…Indeed, the high toxicity level of LPS excludes its use for clinical applications, whereas MPLA represent a low toxicity derivative of LPS available in GMP grade. In combination with IFNγ, MPLA was shown to be an efficient maturation agent for monocyte-derived DC maturation [14][15][16]. As such, we integrated the use of MPLA in the manufacturing process, first by titration using small-scale experiments (24-well culture plates) and by comparing its efficiency to LPS ( Figure 5A).…”

Section: Antigen Loading and Maturation Of Monocyte-derived DC (Day 5-6)mentioning

confidence: 99%

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Development and Optimization of a GMP-Compliant Manufacturing Process for a Personalized Tumor Lysate Dendritic Cell Vaccine

et al. 2020

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With the emergence of immune checkpoint inhibitors and adoptive T-cell therapies, there is a considerable interest in using personalized autologous dendritic cell (DC) vaccines in combination with T cell-targeting immunotherapies to potentially maximize the therapeutic impact of DC vaccines. Here, we describe the development and optimization of a Good Manufacturing Practice (GMP)-compliant manufacturing process based on tumor lysate as a tumor antigen source for the production of an oxidized tumor cell lysate loaded DC (OC-DC) vaccine. The manufacturing process required one day for lysate preparation and six days for OC-DC vaccine production. Tumor lysate production was standardized based on an optimal tumor digestion protocol and the immunogenicity was improved through oxidation using hypochloric acid prior to freeze-thaw cycles resulting in the oxidized tumor cell lysate (OC-L). Next, monocytes were selected using the CliniMACS prodigy closed system and were placed in culture in cell factories in the presence of IL-4 and GM-CSF. Immature DCs were loaded with OC-L and matured using MPLA-IFNγ. After assessing the functionality of the OC-DC cells (IL12p70 secretion and COSTIM assay), the OC-DC vaccine was cryopreserved in multiple doses for single use. Finally, the stability of the formulated doses was tested and validated. We believe this GMP-compliant DC vaccine manufacturing process will facilitate access of patients to personalized DC vaccines, and allow for multi-center clinical trials.

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Section: Discussionsupporting

confidence: 88%

Section: Antigen Loading and Maturation Of Monocyte-derived DC (Day 5-6)mentioning

confidence: 99%

Development and Optimization of a GMP-Compliant Manufacturing Process for a Personalized Tumor Lysate Dendritic Cell Vaccine

et al. 2020

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“…Dauer and colleagues on 2003 described a strategy for differentiation and maturation of monocyte derived DC within only 48-72 h of in vitro culture, called "Fast" protocol, and several other groups have further elaborated on this pioneering work [13][14][15][16][17]. Here we described this method modified from our group to obtain Fast mature autologous DC, loaded with antigens obtained from whole tumor lysate, suitable for the immunotherapy of glioblastoma patients.…”

Section: Discussionmentioning

confidence: 99%

“…Fully mature DC are generally obtained in 7-10 days of culture: differentiation step take place in 5-7 days with GM-CSF and IL4; immature DC are subsequently induced to maturation for 1-3 days with pro-inflammatory stimuli to generate a population of immunogenic mature DC [12]. Several groups have shown that it is possible to obtain mature DC in 2-3 days cell culture [13][14][15][16][17]. The in vitro generation of DC with Fast protocol will shorten the time from the patients' recruiting to DC treatment and therefore be beneficial in the clinical setting: a Fast protocol would result in a simplified processing that minimizes inter-preparations variability and the risk of contamination, also resulting less expensive.…”

Section: Introductionmentioning

confidence: 99%

PGE2 Is Crucial for the Generation of FAST Whole- Tumor-Antigens Loaded Dendritic Cells Suitable for Immunotherapy in Glioblastoma

et al. 2020

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Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5–7 days of differentiation with GM-CSF and IL-4 followed by 2–3 days of activation/maturation. In attempts to shorten the vaccine’s production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination.

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“…To further induce DC activation, mimicking viral infection, type-I interferons have been added to the cocktail [55]. Furthermore recently, utilize of Toll-like receptor (TLR) ligands [56,57] or electroporation with mRNA-encoding proteins that induce DC maturation [58] has been investigated.…”

Section: In Vitro Maturation Of Modcsmentioning

confidence: 99%

In Laboratory Generation and Maturation of Human Monocyte-Derived Dendritic Cells for Cancer Immunotherapy

Mishra¹,

Bharadva²,

Joshi

et al. 2020

Int J App Pharm

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Dendritic cells (DCs) play a critical role in the regulation of adaptive immune responses, furthermore they act as a bridge between the innate and the adaptive immune systems they have been ideal candidates for cell-based immunotherapy of cancers and infections in humans. The first reported trial using DCs in 1995, since they have been used in trials all over the world for several of indications, including cancer and human immunodeficiency virus infection. Generally, for in vitro experiments or for DCs vaccination monocyte-derived dendritic cells (moDCs) were generated from purified monocytes that isolated from peripheral blood by density gradient centrifugation. A variety of methods can be used for enrichment of monocytes for generation of clinical-grade DCs. Herein we summarized up to date understanding of systems and inputs used in procedures to differentiate DCs from blood monocytes in vitro.

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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

Cited by 13 publications

References 52 publications

Development and Optimization of a GMP-Compliant Manufacturing Process for a Personalized Tumor Lysate Dendritic Cell Vaccine

Development and Optimization of a GMP-Compliant Manufacturing Process for a Personalized Tumor Lysate Dendritic Cell Vaccine

PGE2 Is Crucial for the Generation of FAST Whole- Tumor-Antigens Loaded Dendritic Cells Suitable for Immunotherapy in Glioblastoma

In Laboratory Generation and Maturation of Human Monocyte-Derived Dendritic Cells for Cancer Immunotherapy

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