Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets

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...

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Main Authors:	Sköld, Annette E., van Beek, Jasper J. P., Sittig, Simone P., Bakdash, Ghaith, Tel, Jurjen, Schreibelt, Gerty, de Vries, I. Jolanda M.
Format:	Online
Language:	English
Published:	Springer Berlin Heidelberg 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4612318/

id	pubmed-4612318
recordtype	oai_dc
spelling	pubmed-46123182015-10-26 Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets Sköld, Annette E. van Beek, Jasper J. P. Sittig, Simone P. Bakdash, Ghaith Tel, Jurjen Schreibelt, Gerty de Vries, I. Jolanda M. Original Article 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. Springer Berlin Heidelberg 2015-08-15 2015 /pmc/articles/PMC4612318/ /pubmed/26275446 http://dx.doi.org/10.1007/s00262-015-1746-9 Text en © The Author(s) 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Sköld, Annette E. van Beek, Jasper J. P. Sittig, Simone P. Bakdash, Ghaith Tel, Jurjen Schreibelt, Gerty de Vries, I. Jolanda M.
spellingShingle	Sköld, Annette E. van Beek, Jasper J. P. Sittig, Simone P. Bakdash, Ghaith Tel, Jurjen Schreibelt, Gerty de Vries, I. Jolanda M. Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets
author_facet	Sköld, Annette E. van Beek, Jasper J. P. Sittig, Simone P. Bakdash, Ghaith Tel, Jurjen Schreibelt, Gerty de Vries, I. Jolanda M.
author_sort	Sköld, Annette E.
title	Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets
title_short	Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets
title_full	Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets
title_fullStr	Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets
title_full_unstemmed	Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets
title_sort	protamine-stabilized rna as an ex vivo stimulant of primary human dendritic cell subsets
description	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.
publisher	Springer Berlin Heidelberg
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4612318/
_version_	1613490333107093504

Protamine-stabilized RNA as an ex vivo stimulant of primary human dendritic cell subsets

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