Polyinosinic polycytidylic acid prevents efficient antigen expression after mRNA electroporation of clinical grade dendritic cells.
Tumor-derived peptides are used frequently as antigen (Ag) source in dendritic cell (DC) therapy in cancer patients. An alternative is to load DC with tumor-associated Ag (TAA)-encoding RNA. RNA-loading obviates prior knowledge of CTL and Th epitopes in the Ag. Multiple epitopes for many HLA alleles (both MHC class I and class II) are encoded by the RNA and loading is independent of the patient's HLA make-up. Herein, we determined the optimal conditions for mRNA-electroporation of monocyte-derived DC for clinical application in relation to different maturation cocktails. The data demonstrate that TAA carcinoembryonic antigen, gp100 and tyrosinase are expressed already 30 min after electroporation with the encoding mRNA. Moreover, gp100-specific CTL are activated by gp100 mRNA-electroporated DC. Importantly, we show here that the presence of polyinosinic-polycytidylic acid [poly(I:C)] in the maturation cocktail prevents effective protein expression of the electroporated mRNA as well as subsequent CTL recognition. This effect of poly(I:C) correlates with the induction of IFN-induced genes and innate anti-viral effector molecules in DC. Together these data show that electroporation of mature DC with TAA-encoding mRNA is attractive for use in DC vaccination protocols in cancer patients, but protein expression should be tested for each maturation cocktail.
Schuurhuis
Danita H
DH
Department of Tumor Immunology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands. d.beekmanschuurhuis@genmab.com
Lesterhuis
W Joost
WJ
Kramer
Matthijs
M
Looman
Maaike G M
MG
van Hout-Kuijer
Maaike
M
Schreibelt
Gerty
G
Boullart
A C Inge
AC
Aarntzen
Erik H J G
EH
Benitez-Ribas
Daniel
D
Figdor
Carl G
CG
Punt
Cornelis J A
CJ
de Vries
I Jolanda M
IJ
Adema
Gosse J
GJ
D017951:Antigen Presentation / Q000187:drug effects* / Q000276:immunology; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D002272:Carcinoembryonic Antigen / Q000235:genetics / Q000276:immunology; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology* / Q000637:transplantation; D018274:Electroporation; D006801:Humans; D007369:Interferon Inducers / Q000494:pharmacology*; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000276:immunology; D011070:Poly I-C / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology; D058951:gp100 Melanoma Antigen
D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D007369:Interferon Inducers; D008562:Membrane Glycoproteins; C548637:PMEL protein, human; D012333:RNA, Messenger; D058951:gp100 Melanoma Antigen; D007371:Interferon-gamma; D014442:Monophenol Monooxygenase; D011070:Poly I-C