PMID- 11730854
OWN - NLM
STAT- MEDLINE
DCOM- 20020201
LR  - 20210103
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 259
IP  - 1-2
DP  - 2002 Jan 1
TI  - mRNA-based electrotransfection of human dendritic cells and induction of 
      cytotoxic T lymphocyte responses against the telomerase catalytic subunit 
      (hTERT).
PG  - 191-203
AB  - Dendritic cells (DCs) are recognised as the most potent antigen-presenting cells 
      for induction of cellular immune responses, and vaccination with DCs pulsed with 
      antigens has emerged as a promising strategy for generating protective immunity 
      in mammals. We have developed a transfection method that uses in vitro 
      synthesised mRNA and square-wave electroporation for transient expression in DCs 
      and other cell types. The method is highly efficient and produces almost complete 
      transfection of cells in culture. When using mRNA encoding the enhanced green 
      fluorescence protein (EGFP), highest expression in DCs occurred on the second day 
      after transfection and produced a 76-fold increase in mean fluorescence above 
      background. High levels of expression were maintained for at least 5 days 
      post-transfection. In comparison, square-wave electroporation of DCs with EGFP 
      plasmid DNA yielded 15% transfected cells and a 28-fold increase of mean 
      fluorescence. DCs transfected with mRNA encoding the telomerase catalytic subunit 
      (hTERT) acquired strong telomerase activity and were capable of eliciting a 
      hTERT-specific cytotoxic T lymphocyte (CTL) response in vitro.
FAU - Saeboe-Larssen, Stein
AU  - Saeboe-Larssen S
AD  - Section for Immunotherapy, Department of Immunology, The Norwegian Radium 
      Hospital, Cancer Research Institute, University of Oslo, N-0310 Montebello, Oslo, 
      Norway. stein.saboe-larssen@samfunnsmed.uio.no
FAU - Fossberg, Ellen
AU  - Fossberg E
FAU - Gaudernack, Gustav
AU  - Gaudernack G
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (RNA, Messenger)
RN  - EC 2.7.7.49 (Telomerase)
SB  - IM
MH  - Antigen Presentation/*genetics/*immunology
MH  - Catalytic Domain/genetics/immunology
MH  - Cells, Cultured
MH  - Cytotoxicity, Immunologic
MH  - DNA-Binding Proteins
MH  - Dendritic Cells/*immunology
MH  - Electroporation
MH  - Humans
MH  - RNA, Messenger/genetics/immunology
MH  - T-Lymphocytes, Cytotoxic/*immunology
MH  - Telomerase/*genetics/*immunology
MH  - Transfection/*methods
EDAT- 2001/12/04 10:00
MHDA- 2002/02/02 10:01
CRDT- 2001/12/04 10:00
PHST- 2001/12/04 10:00 [pubmed]
PHST- 2002/02/02 10:01 [medline]
PHST- 2001/12/04 10:00 [entrez]
AID - S0022175901005063 [pii]
AID - 10.1016/s0022-1759(01)00506-3 [doi]
PST - ppublish
SO  - J Immunol Methods. 2002 Jan 1;259(1-2):191-203. doi: 
      10.1016/s0022-1759(01)00506-3.