PMID- 26275446
OWN - NLM
STAT- MEDLINE
DCOM- 20160202
LR  - 20201226
IS  - 1432-0851 (Electronic)
IS  - 0340-7004 (Print)
IS  - 0340-7004 (Linking)
VI  - 64
IP  - 11
DP  - 2015 Nov
TI  - Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell 
      subsets.
PG  - 1461-73
LID - 10.1007/s00262-015-1746-9 [doi]
AB  - Dendritic cells (DCs) are key in connecting innate and adaptive immunity. Their 
      potential in inducing specific immune responses has made them interesting targets 
      for immunotherapeutic approaches. Our research group was the first to exploit the 
      naturally occurring myeloid DCs (mDCs) and plasmacytoid DCs (pDCs) in therapeutic 
      vaccination trials against melanoma. To develop primary DC subsets as an optimal 
      vaccine, the identification of a clinically applicable adjuvant activating both 
      subsets is required. Although the expression of pathogen recognition receptors 
      differs distinctly between the DC subsets, both pDCs and mDCs can respond to 
      single-stranded RNA (ssRNA) via Toll-like receptors 7 and 8, respectively. Since 
      ssRNA is easily degraded by RNases, we stabilized anionic RNA by complexing it 
      with the positively charged protein protamine. This leads to the formation of 
      protamine-RNA complexes with varying features depending on ionic content. We 
      subsequently investigated the immunostimulatory effect of complexes that formed 
      various salt concentrations on purified DC subsets. Both mDCs and pDCs 
      upregulated maturation markers and produced pro-inflammatory cytokines in a 
      dose-dependent way to the protamine-RNA complexes. This was dependent on 
      endosomal acidification and correlated partly with the uptake of protamine-RNA 
      complexes. Furthermore, both DC subsets induced T cell proliferation and IFN 
      gamma secretion in a beneficial ratio to IL-10. These results indicate that 
      protamine-RNA complexes can be used to stimulate human mDC and pDC ex vivo for 
      use in immunotherapeutic settings.
FAU - Skold, Annette E
AU  - Skold AE
AUID- ORCID: 0000-0002-5309-8860
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands.
AD  - Department of Oncology-Pathology, Karolinska University Hospital Solna, 
      Karolinska Institutet, Stockholm, Sweden.
FAU - van Beek, Jasper J P
AU  - van Beek JJ
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands.
FAU - Sittig, Simone P
AU  - Sittig SP
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands.
FAU - Bakdash, Ghaith
AU  - Bakdash G
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands.
FAU - Tel, Jurjen
AU  - Tel J
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands.
FAU - Schreibelt, Gerty
AU  - Schreibelt G
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands.
FAU - de Vries, I Jolanda M
AU  - de Vries IJ
AD  - Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Centre, Nijmegen, The Netherlands. 
      Jolanda.deVries@radboudumc.nl.
AD  - Department of Medical Oncology, Radboud University Medical Centre, Geert 
      Grooteplein 26, Route 278, 6525 GA, Nijmegen, The Netherlands. 
      Jolanda.deVries@radboudumc.nl.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150815
PL  - Germany
TA  - Cancer Immunol Immunother
JT  - Cancer immunology, immunotherapy : CII
JID - 8605732
RN  - 0 (Adjuvants, Immunologic)
RN  - 0 (Cytokines)
RN  - 0 (Protamines)
RN  - 0 (Toll-Like Receptors)
RN  - 451W47IQ8X (Sodium Chloride)
RN  - 63231-63-0 (RNA)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Adjuvants, Immunologic/*pharmacology
MH  - Cells, Cultured
MH  - Cytokines/metabolism
MH  - Dendritic Cells/classification/*immunology
MH  - Dose-Response Relationship, Drug
MH  - Endosomes/physiology
MH  - Humans
MH  - Interferon-gamma/biosynthesis
MH  - Lymphocyte Activation
MH  - Protamines/*pharmacology
MH  - RNA/*pharmacology
MH  - RNA Stability
MH  - Sodium Chloride/pharmacology
MH  - Toll-Like Receptors/physiology
PMC - PMC4612318
OTO - NOTNLM
OT  - Adjuvants
OT  - GMP
OT  - Immunotherapy
OT  - TLR
OT  - mDC
OT  - pDC
COIS- The authors declare no conflicts of interest.
EDAT- 2015/08/16 06:00
MHDA- 2016/02/03 06:00
PMCR- 2015/08/15
CRDT- 2015/08/16 06:00
PHST- 2014/10/31 00:00 [received]
PHST- 2015/07/21 00:00 [accepted]
PHST- 2015/08/16 06:00 [entrez]
PHST- 2015/08/16 06:00 [pubmed]
PHST- 2016/02/03 06:00 [medline]
PHST- 2015/08/15 00:00 [pmc-release]
AID - 10.1007/s00262-015-1746-9 [pii]
AID - 1746 [pii]
AID - 10.1007/s00262-015-1746-9 [doi]
PST - ppublish
SO  - Cancer Immunol Immunother. 2015 Nov;64(11):1461-73. doi: 
      10.1007/s00262-015-1746-9. Epub 2015 Aug 15.