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Restricted cytokine production from mouse peritoneal macrophages in culture in spite of extensive uptake of plasmid DNA
Author(s) -
Yasuda Kei,
Kawano Hiroki,
Yamane Ikuko,
Ogawa Yoshiyuki,
Yoshinaga Takaharu,
Nishikawa Makiya,
Takakura Yoshinobu
Publication year - 2004
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2004.01814.x
Subject(s) - plasmid , cytokine , biology , dna , microbiology and biotechnology , immunology , genetics
Summary The production of inflammatory cytokines from macrophages (Mφ), 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 Mφ activation, following stimulation with naked pDNA, based on the production of cytokines from cell lines (RAW264.7 and J774A1) and peritoneal Mφs in primary culture. The Mφ cell lines RAW264.7 and J774A1 produced a significant amount of tumour necrosis factor‐α (TNF‐α) upon stimulation with naked pDNA and this response required endosomal acidification. On the other hand, peritoneal Mφs (both resident and elicited) in primary culture did not secrete TNF‐α 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 Mφs showed a higher uptake of pDNA than the RAW264.7 and J774A1 cell lines. These findings have important implications for Mφ 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 Mφ cell lines.