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.
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.
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.
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.
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http://dx.doi.org/10.1016/j.jcyt.2018.06.006 | DOI Listing |
Nat Rev Gastroenterol Hepatol
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Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Ministerio de Sanidad, Madrid, Spain.
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January 2025
Department of Chemistry, Yazd University, Yazd, Iran.
A new humic acid-based nanomagnetic copper(II) composite was prepared and used as an eco-friendly recoverable catalyst for synthesizing 1,4-disubstituted 1,2,3-triazoles. The synthesis was done via the three-component click reaction of alkyl halide, sodium azide, and terminal alkyne with good to excellent yield. A simple magnetic copper acetate composite, FeO@HA-Cu(OAc), was prepared using humic acid and characterized by SEM, TEM, XRD, EDX, EDS-mapping, VSM, TGA, AAS, and FT-IR.
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January 2025
Department of Chemistry, Faculty of Science, Arak University, Arak, 38481-77584, Iran.
In this study, a novel hybrid nanostructure consisting of acid-decorated chitosan and magnetic AlFeO nanoparticles was fabricated. The acid-decorated chitosan provided a stable and biocompatible matrix for the magnetic AlFeO nanoparticles. Various techniques including Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction patterns (XRD), thermogravimetric analysis (TGA), vibrating sample magnetometry (VSM), specific surface area (BET), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) were used to characterize and confirm the successful synthesis of the hybrid nanostructure.
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January 2025
College of Life Sciences, Northwest A&F University, Yangling, 712100, China.
The contamination of Chinese medicinal materials with cadmium (Cd) is a pressing global issue that poses significant risks to human health. The beneficial effects of selenium (Se) have been established in improving plant growth and reducing Cd accumulation in plant under Cd stress. This study employed soil cultivation experiments to investigate the remediation effects of exogenous Se (0, 0.
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January 2025
Centre for Informatics and Systems of the University of Coimbra, Department of Informatics Engineering, University of Coimbra, Coimbra, Portugal.
Assessing cognitive load using pupillography frequency features presents a persistent challenge due to the lack of consensus on optimal frequency limits. This study aims to address this challenge by exploring pupillography frequency bands and seeking clarity in defining the most effective ranges for cognitive load assessment. From a controlled experiment involving 21 programmers performing software bug inspection, our study pinpoints the optimal low-frequency (0.
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