Optimized Protocols for siRNA Delivery into Monocytes and Dendritic Cells

RNA interference technology is an ideal strategy to elucidate the mechanisms associated with humanCD34+hematopoietic stem cell differentiation into dendritic cells. Simple manipulations in vitro can unequivocally yield alloreactive or tolerogenic populations, suggesting key implications of biochemical players that might emerge as therapeutic targets for cancer or graft-versus-host disease. To knockdown proteins typically involved in the biology of dendritic cells, we employed an siRNA delivery system based on the cationic liposome DOTAP as the carrier. Freshly-isolatedCD34+cells were transfected with siRNA for cathepsin S with negligible cytotoxicity and transfection rates (>60%) comparable to the efficiency shown by lentiviral vectors. Further, cathepsin S knockdown was performed during both cell commitment and through the entire 14-day differentiation process with repeated transfection rounds that had no effect per se on cell development. Tested in parallel, other commercially-available chemical reagents failed to meet acceptable standards. In addition to safe and practical handling, a direct advantage of DOTAP over viral-mediated techniques is that transient silencing effects can be dynamically appraised through the recovery of targeted proteins. Thus, our findings identify DOTAP as an excellent reagent for gene silencing in resting and differentiatingCD34+cells, suggesting a potential for applications in related preclinical models.

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Small interfering RNA (siRNA) delivery into monocyte-derived dendritic cells by electroporation

Journal of Immunological Methods ◽

10.1016/j.jim.2006.01.021 ◽

2006 ◽

Vol 311 (1-2) ◽

pp. 139-152 ◽

Cited By ~ 41

Author(s):

Alexander T. Prechtel ◽

Nadine M. Turza ◽

Alexandros A. Theodoridis ◽

Mirko Kummer ◽

Alexander Steinkasserer

Keyword(s):

Dendritic Cells ◽

Small Interfering Rna ◽

Sirna Delivery ◽

Interfering Rna

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Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation☆

Journal of Controlled Release ◽

10.1016/j.jconrel.2010.01.012 ◽

2010 ◽

Vol 143 (3) ◽

pp. 311-317 ◽

Cited By ~ 100

Author(s):

Hidetaka Akita ◽

Kentaro Kogure ◽

Rumiko Moriguchi ◽

Yoshio Nakamura ◽

Tomoko Higashi ◽

...

Keyword(s):

Dendritic Cells ◽

Cancer Vaccines ◽

Ex Vivo ◽

Sirna Delivery ◽

Endosomal Escape

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Reprint of: Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation

Journal of Controlled Release ◽

10.1016/j.jconrel.2010.08.023 ◽

2011 ◽

Vol 149 (1) ◽

pp. 58-64 ◽

Cited By ~ 20

Author(s):

Hidetaka Akita ◽

Kentaro Kogure ◽

Rumiko Moriguchi ◽

Yoshio Nakamura ◽

Tomoko Higashi ◽

...

Keyword(s):

Dendritic Cells ◽

Cancer Vaccines ◽

Ex Vivo ◽

Sirna Delivery ◽

Endosomal Escape

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Small interfering RNA (siRNA) delivery into murine bone marrow-derived dendritic cells by electroporation

Journal of Immunological Methods ◽

10.1016/j.jim.2008.04.004 ◽

2008 ◽

Vol 337 (1) ◽

pp. 71-77 ◽

Cited By ~ 27

Author(s):

Jonathan Jantsch ◽

Nadine Turza ◽

Melanie Volke ◽

Kai-Uwe Eckardt ◽

Michael Hensel ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Small Interfering Rna ◽

Sirna Delivery ◽

Murine Bone Marrow ◽

Interfering Rna

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A novel in vivo siRNA delivery system specifically targeting dendritic cells and silencing CD40 genes for immunomodulation

Blood ◽

10.1182/blood-2008-04-151191 ◽

2009 ◽

Vol 113 (12) ◽

pp. 2646-2654 ◽

Cited By ~ 68

Author(s):

Xiufen Zheng ◽

Costin Vladau ◽

Xusheng Zhang ◽

Motohiko Suzuki ◽

Thomas E. Ichim ◽

...

Keyword(s):

Dendritic Cells ◽

Delivery System ◽

Immune Modulation ◽

Sirna Delivery ◽

Specific Cell ◽

Interfering Rna ◽

In Vivo Administration ◽

Sirna Delivery System

Abstract Translation of small interfering RNA (siRNA)–based approaches into practical therapeutics is limited because of lack of an effective and cell-specific delivery system. Herein, we present a new method of selectively delivering siRNA to dendritic cells (DCs) in vivo using CD40 siRNA-containing immunoliposomes (siILs) that were decorated with DC-specific DEC-205 mAb. Administration of CD40 siILs resulted in DC-specific cell targeting in vitro and in vivo. On treatment with CD40 siILs, the expression of CD40 in DCs, as well allostimulatory activity was inhibited. In vivo administration resulted in selective siRNA uptake into immune organs and functional immune modulation as assessed using a model antigen. In conclusion, this is the first demonstration of DC-specific siRNA delivery and gene silencing in vivo, which highlights the potential of DC-mediated immune modulation and the feasibility of siRNA-based clinical therapy.

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Dendritic cells of the human epidermis: immuno-electron microscopic studies of la antigen reactivity

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100084090 ◽

1978 ◽

Vol 36 (2) ◽

pp. 644-645

Author(s):

G. Rowden ◽

M. G. Lewis ◽

T. M. Phillips

Keyword(s):

Dendritic Cells ◽

Langerhans Cells ◽

Cell Types ◽

Cell Type ◽

Electron Microscopic ◽

La Antigen ◽

Microscopic Studies ◽

A Cell ◽

C3 Receptors ◽

Antigen Reactivity

Langerhans cells of mammalian stratified squamous epithelial have proven to be an enigma since their discovery in 1868. These dendritic suprabasal cells have been considered as related to melanocytes either as effete cells, or as post divisional products. Although grafting experiments seemed to demonstrate the independence of the cell types, much confusion still exists. The presence in the epidermis of a cell type with morphological features seemingly shared by melanocytes and Langerhans cells has been especially troublesome. This so called "indeterminate", or " -dendritic cell" lacks both Langerhans cells granules and melanosomes, yet it is clearly not a keratinocyte. Suggestions have been made that it is related to either Langerhans cells or melanocyte. Recent studies have unequivocally demonstrated that Langerhans cells are independent cells with immune function. They display Fc and C3 receptors on their surface as well as la (immune region associated) antigens.

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