title: Choose your models wisely: how different murine bone marrow-derived dendritic cell protocols influence the success of nanoparticulate vaccines in vitro.
abstract: Dendritic cell (DC)-based cancer vaccination has shown great potential in cancer immunotherapy. As a result, novel nanoparticles aiming to load DCs with tumor antigens are being developed and evaluated in vitro. For this, murine bone marrow-derived DCs (BM-DCs) are most commonly used as model DCs. However, many different protocols exist to generate these cells. Therefore, we investigated to what extent different BM-DC culture protocols impact on the immunobiology of the cells, as well as their response to particulate antigens. We evaluated 4 different BM-DC protocols with 2 main variables: bovine serum and cytokine combinations. Our results show distinct differences in yield, phenotypical maturation status and the production of immune stimulatory and immune suppressive cytokines by the different BM-DCs. Importantly, we demonstrate that the antigen-loading of these different BM-DCs via transfection with mRNA lipoplexes results in large differences in transfection efficiency as well as in the capacity of mRNA-transfected BM-DCs to stimulate antigen-specific T cells. Thus, it is clear that the BM-DC model can have significant confounding effects on the evaluation of novel nanoparticulate vaccines. To take this into account when testing novel particulate antigen-delivery systems in BM-DC models, we propose to (1) perform a thorough immunological characterization of the BM-DCs and to (2) not only judge a particle's potential for cancer vaccination based on transfection efficiency, but also to include an evaluation of functional end-points such as T cell activation.
authors: Lab for General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
authors: Heleen
authors: H
authors: Dewitte
authors: Lab for General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
authors: Rein
authors: R
authors: Verbeke
authors: Laboratory of Molecular and Cellular Therapy, Department of Immunology-Physiology, Medical School of the Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1050 Jette, Belgium.
authors: Karine
authors: K
authors: Breckpot
authors: Inflammation Research Center (IRC), VIB, Technologiepark 927, 9052 Ghent, Belgium; Molecular Mouse Genetics Unit, Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Ghent, Belgium.
authors: Roosmarijn E
authors: RE
authors: Vandenbroucke
authors: Inflammation Research Center (IRC), VIB, Technologiepark 927, 9052 Ghent, Belgium; Molecular Mouse Genetics Unit, Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Ghent, Belgium.
authors: Claude
authors: C
authors: Libert
authors: Lab for General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium. Electronic address: Stefaan.DeSmedt@UGent.be.
authors: Stefaan C
authors: SC
authors: De Smedt
authors: Lab for General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
authors: Ine
authors: I
authors: Lentacker
mesh_terms: D000818:Animals; D000941:Antigens / Q000235:genetics*; D051940:B7-2 Antigen / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology; D019013:CD40 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D005802:Genes, MHC Class II / Q000276:immunology; D049452:Green Fluorescent Proteins / Q000235:genetics; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000008:administration & dosage*; D010047:Ovalbumin / Q000235:genetics; D010641:Phenotype; D012333:RNA, Messenger / Q000008:administration & dosage*; D014612:Vaccines / Q000008:administration & dosage*
chemical_list: D000941:Antigens; D051940:B7-2 Antigen; D019013:CD40 Antigens; D016207:Cytokines; D008081:Liposomes; D012333:RNA, Messenger; D014612:Vaccines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D010047:Ovalbumin; D008156:Luciferases
keywords: Cancer vaccination; CholEsteryl BODIPY® FL C12 (PubChem CID: 70682631); DOPE (PubChem CID: 9546757); DOTAP (PubChem CID: 6437371); Dendritic cell; Nanoparticle; SIINFEKL (PubChem CID: 71311993); mRNA