Design of bifunctional siRNAs: Combining immunostimulation and gene-silencing in one single siRNA molecule

Active suppression of T lymphocyte activation can limit the efficacy of immune surveillance and immunotherapy. Here we have explored the possibility of designing bifunctional small interfering RNAs (siRNAs) capable of inducing innate immunity through Toll-like receptors and simultaneously inhibiting...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2007-01, Vol.352 (3), p.642-649
Main Authors: Furset, Gro, Sioud, Mouldy
Format: Article
Language:English
Subjects:
Cells, Cultured
Dendritic cells
Gene Silencing - immunology
Genetic Engineering - methods
Humans
Immunization - methods
Innate immunity
Interferon pathway
Interleukin-10 - immunology
Leukocytes, Mononuclear - immunology
Protein kinase R
RNA interference
RNA, Small Interfering - genetics
RNA, Small Interfering - immunology
siRNAs
Toll-like receptors
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Summary:Active suppression of T lymphocyte activation can limit the efficacy of immune surveillance and immunotherapy. Here we have explored the possibility of designing bifunctional small interfering RNAs (siRNAs) capable of inducing innate immunity through Toll-like receptors and simultaneously inhibiting the expression of immunosuppressive factors. Using interleukin (IL) 10 as a model, we found that liposomal delivery of IL10 siRNAs could efficiently activate the expression of cytokines (e.g. TNF-α, IL6, and IL12) and interferons (e.g. IFN-α) in peripheral blood mononuclear cells (PBMCs) and immature monocyte-derived dendritic cells (iMoDCs). Moreover, the designed siRNAs inhibited IL10 gene expression. Transfection of iMoDCs with either chemically or in vitro transcribed IL10 siRNAs induced their differentiation into mature MoDCs (mMoDCs) characterized by the expression of costimulatory molecules CD80/CD86 and the chemokine receptor CCR7. Lipid delivery of either chemically synthesized or T7-transcribed immunostimulatory siRNAs induced cytokine production. However, in contrast to chemically synthesized siRNAs, electroporation of in vitro transcribed siRNAs also induced cytokine production in iMoDCs. Interestingly, IL10 siRNA-transfected iMoDCs were capable for enhancing the response of allogeneic T cells, providing support for the rational design of bifunctional siRNAs as immune modulating therapy.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2006.11.059