PMID- 17626798
OWN - NLM
STAT- MEDLINE
DCOM- 20071218
LR  - 20171116
IS  - 0741-5400 (Print)
IS  - 0741-5400 (Linking)
VI  - 82
IP  - 4
DP  - 2007 Oct
TI  - High transfection efficiency, gene expression, and viability of monocyte-derived 
      human dendritic cells after nonviral gene transfer.
PG  - 849-60
AB  - Dendritic cells (DCs) are bone marrow-originated, professional antigen-capturing 
      cells and APCs, which can function as vaccine carriers. Although efficient 
      transfection of human DCs has been achieved with viral vectors, viral gene 
      products may influence cellular functions. In contrast, nonviral methods have 
      generally resulted in inefficient gene transfer, low levels of gene expression, 
      and/or low cell viability. Monocyte-derived DCs are the most common source of DCs 
      for in vitro studies and for in vivo applications. We hypothesized that reduction 
      of the time to generate immature DCs (iDCs) might result in higher viability 
      after transfection. Therefore, we established a protocol to generate human iDCs 
      from CD14(+) monocytes within 3 days. These "fast" iDCs were phenotypically and 
      functionally indistinguishable from conventional iDCs, showing high endocytic 
      ability and low antigen-presenting capacity. Furthermore, the fast iDCs matured 
      normally and had similar antigen-presenting capacity to conventional mature DCs. 
      To optimize transfection of iDCs, we compared nonviral transfection of plasmid 
      DNA and in vitro-transcribed (IVT) RNA with transfection reagents, 
      electroporation, and nucleofection. Nucleofection of IVT RNA with the X1 program 
      of an Amaxa Co. Nucleofector resulted in the most efficient transfection, with an 
      average of 93% transfected iDCs, excellent long-term viability, and strong 
      protein expression. Furthermore, the IVT RNA-transfected iDCs retained all 
      phenotypic and functional characteristics of iDCs. This method is applicable to 
      most purposes, including in vitro functional assays, in vivo DC immunotherapy, 
      and DC-based vaccines.
FAU - Landi, Abdolamir
AU  - Landi A
AD  - Vaccine and Infectious Disease Organization, University of Saskatchewan, 120 
      Veterinary Rd., Saskatoon, SK, S7N 5E3, Canada.
FAU - Babiuk, Lorne A
AU  - Babiuk LA
FAU - van Drunen Littel-van den Hurk, Sylvia
AU  - van Drunen Littel-van den Hurk S
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070712
PL  - England
TA  - J Leukoc Biol
JT  - Journal of leukocyte biology
JID - 8405628
RN  - 0 (Lipopolysaccharide Receptors)
RN  - 0 (Vaccines)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Antigen Presentation/*immunology
MH  - Cell Survival/immunology
MH  - Cells, Cultured
MH  - *Dendritic Cells/cytology/immunology
MH  - Electroporation
MH  - Endocytosis/immunology
MH  - *Gene Expression
MH  - Humans
MH  - *Immunotherapy, Adoptive
MH  - Lipopolysaccharide Receptors/immunology
MH  - *Monocytes/cytology/immunology
MH  - Plasmids/immunology
MH  - RNA
MH  - Time Factors
MH  - *Transfection
MH  - Vaccines/immunology
EDAT- 2007/07/14 09:00
MHDA- 2007/12/19 09:00
CRDT- 2007/07/14 09:00
PHST- 2007/07/14 09:00 [pubmed]
PHST- 2007/12/19 09:00 [medline]
PHST- 2007/07/14 09:00 [entrez]
AID - jlb.0906561 [pii]
AID - 10.1189/jlb.0906561 [doi]
PST - ppublish
SO  - J Leukoc Biol. 2007 Oct;82(4):849-60. doi: 10.1189/jlb.0906561. Epub 2007 Jul 12.