title: Electroporated Antigen-Encoding mRNA Is Not a Danger Signal to Human Mature Monocyte-Derived Dendritic Cells.
abstract: For therapeutic cancer vaccination, the adoptive transfer of mRNA-electroporated dendritic cells (DCs) is frequently performed, usually with monocyte-derived, cytokine-matured DCs (moDCs). However, DCs are rich in danger-sensing receptors which could recognize the exogenously delivered mRNA and induce DC activation, hence influencing the DCs' immunogenicity. Therefore, we examined whether electroporation of mRNA with a proper cap and a poly-A tail of at least 64 adenosines had any influence on cocktail-matured moDCs. We used 16 different RNAs, encoding tumor antigens (MelanA, NRAS, BRAF, GNAQ, GNA11, and WT1), and variants thereof. None of those RNAs induced changes in the expression of CD25, CD40, CD83, CD86, and CD70 or the secretion of the cytokines IL-8, IL-6, and TNFα of more than 1.5-fold compared to the control condition, while an mRNA encoding an NF-κB-activation protein as positive control induced massive secretion of the cytokines. To determine whether mRNA electroporation had any effect on the whole transcriptome of the DCs, we performed microarray analyses of DCs of 6 different donors. None of 60,000 probes was significantly different between mock-electroporated DCs and MelanA-transfected DCs. Hence, we conclude that no transcriptional programs were induced within cocktail-matured DCs by electroporation of single tumor-antigen-encoding mRNAs.
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Hoyer
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany; Department of Genetics, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Kerstin F
authors.initials: KF
authors.lastname: Gerer
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Isabell A
authors.initials: IA
authors.lastname: Pfeiffer
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany; Department of Genetics, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Prommersberger
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany; Department of Genetics, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Höfflin
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Tanushree
authors.initials: T
authors.lastname: Jaitly
authors.affiliation: Department of Oncology, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Beltrame
authors.affiliation: Department of Neurosciences, Psychology, Drug Research and Child Health, University of Florence, Florence, Italy.
authors.forename: Duccio
authors.initials: D
authors.lastname: Cavalieri
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Julio
authors.initials: J
authors.lastname: Vera
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
mesh_terms: D019496:Cancer Vaccines / Q000276:immunology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000502:physiology* / Q000637:transplantation; D018274:Electroporation; D020869:Gene Expression Profiling; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D058965:MART-1 Antigen / Q000235:genetics / Q000378:metabolism*; D046228:Microarray Analysis; D009000:Monocytes / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: D019496:Cancer Vaccines; D016207:Cytokines; D058965:MART-1 Antigen; D012333:RNA, Messenger
references.citation: Am J Clin Exp Immunol. 2013 Oct 16;2(3):195-207
references.pmid: 24179728
references.citation: J Immunol. 2005 Mar 1;174(5):3087-97
references.pmid: 15728524
references.citation: J Biol Chem. 2002 Apr 12;277(15):12689-96
references.pmid: 11821398
references.citation: Eur J Immunol. 2007 Nov;37 Suppl 1:S53-60
references.pmid: 17972346
references.citation: Immunity. 2013 Jul 25;39(1):38-48
references.pmid: 23890062
references.citation: J Biol Chem. 2004 Mar 26;279(13):12542-50
references.pmid: 14729660
references.citation: Front Cell Infect Microbiol. 2013;3:37
references.pmid: 23908972
references.citation: Eur J Immunol. 2009 Sep;39(9):2537-47
references.pmid: 19662634
references.citation: Eur J Immunol. 2014 Nov;44(11):3413-28
references.pmid: 25100611
references.citation: Science. 2004 Mar 5;303(5663):1526-9
references.pmid: 14976262
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5598-603
references.pmid: 15034168
references.citation: Mol Ther. 2004 May;9(5):757-64
references.pmid: 15120337
references.citation: Clin Cancer Res. 2013 Oct 15;19(20 Suppl):S4-98
references.pmid: 24045178
references.citation: J Gene Med. 2005 Apr;7(4):452-65
references.pmid: 15515120
references.citation: Gene Ther. 2005 May;12(9):772-82
references.pmid: 15750615
references.citation: Cancer Immunol Immunother. 2015 Nov;64(11):1461-73
references.pmid: 26275446
references.citation: Cancer Immunol Immunother. 2014 Sep;63(9):959-67
references.pmid: 24878889
references.citation: Science. 2004 Mar 5;303(5663):1529-31
references.pmid: 14976261
references.citation: Eur J Immunol. 2010 Aug;40(8):2123-30
references.pmid: 20853498
references.citation: Int Rev Immunol. 2011 Feb;30(1):16-34
references.pmid: 21235323
references.citation: Nucleic Acids Res. 2015 Apr 20;43(7):e47
references.pmid: 25605792
references.citation: J Immunother. 2008 Oct;31(8):731-41
references.pmid: 18779746
references.citation: J Immunol. 2004 Jun 1;172(11):6649-57
references.pmid: 15153480
references.citation: Curr Top Microbiol Immunol. 2009;335:107-21
references.pmid: 19802562
references.citation: J Exp Med. 1999 Mar 1;189(5):821-9
references.pmid: 10049946
references.citation: Curr Opin Immunol. 2011 Jun;23(3):399-406
references.pmid: 21497074
references.citation: Expert Rev Vaccines. 2015 Feb;14(2):161-76
references.pmid: 25196947
references.citation: Cancer Immunol Immunother. 2008 Apr;57(4):467-77
references.pmid: 17768622
references.citation: J Immunother. 2007 Sep;30(6):663-74
references.pmid: 17667530
references.citation: Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
references.pmid: 12824337