Document Detail

Restricted cytokine production from mouse peritoneal macrophages in culture in spite of extensive uptake of plasmid DNA.
MedLine Citation:
PMID:  15009428     Owner:  NLM     Status:  MEDLINE    
The production of inflammatory cytokines from macrophages (Mphi), upon stimulation with plasmid DNA (pDNA) containing CpG motifs, is a critical process for DNA-based therapies such as DNA vaccination and gene therapy. We compared Mphi activation, following stimulation with naked pDNA, based on the production of cytokines from cell lines (RAW264.7 and J774A1) and peritoneal Mphis in primary culture. The Mphi cell lines RAW264.7 and J774A1 produced a significant amount of tumour necrosis factor-alpha (TNF-alpha) upon stimulation with naked pDNA and this response required endosomal acidification. On the other hand, peritoneal Mphis (both resident and elicited) in primary culture did not secrete TNF-alpha or interleukin-6, although they contain the mRNA of toll-like receptor-9 (TLR-9) and are able to respond to CpG oligodeoxynucleotides. This unresponsiveness was not a result of impaired cellular uptake of pDNA because the primary cultured Mphis showed a higher uptake of pDNA than the RAW264.7 and J774A1 cell lines. These findings have important implications for Mphi activation by naked pDNA as it has been generally assumed that pDNA that contains CpG motifs is a potent agent for inducing inflammatory cytokines in vivo, based on evidence from in vitro studies using Mphi cell lines.
Kei Yasuda; Hiroki Kawano; Ikuko Yamane; Yoshiyuki Ogawa; Takaharu Yoshinaga; Makiya Nishikawa; Yoshinobu Takakura
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Immunology     Volume:  111     ISSN:  0019-2805     ISO Abbreviation:  Immunology     Publication Date:  2004 Mar 
Date Detail:
Created Date:  2004-03-10     Completed Date:  2004-05-27     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0374672     Medline TA:  Immunology     Country:  England    
Other Details:
Languages:  eng     Pagination:  282-90     Citation Subset:  IM    
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University, 46-29 Yoshida-shimoadachi-cho, Sakyo-ku, Kyoto 606-8501, Japan.
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MeSH Terms
Cell Line
Cells, Cultured
Cytokines / biosynthesis*
DNA, Circular / immunology
DNA, Single-Stranded / immunology
DNA-Binding Proteins / immunology
Enzyme-Linked Immunosorbent Assay / methods
Interleukin-6 / biosynthesis
Macrophage Activation / immunology*
Macrophages, Peritoneal / immunology*
Mice, Inbred ICR
Plasmids / immunology*
RNA, Messenger / analysis
Receptors, Cell Surface / immunology
Reverse Transcriptase Polymerase Chain Reaction / methods
Toll-Like Receptor 9
Tumor Necrosis Factor-alpha / biosynthesis
Reg. No./Substance:
0/Cytokines; 0/DNA, Circular; 0/DNA, Single-Stranded; 0/DNA-Binding Proteins; 0/Interleukin-6; 0/RNA, Messenger; 0/Receptors, Cell Surface; 0/Tlr9 protein, mouse; 0/Toll-Like Receptor 9; 0/Tumor Necrosis Factor-alpha

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