title: Lipopolysaccharide signal transduction in oral keratinocytes--involvement of CD59 but not CD14.
abstract: There are a variety of dermal and mucosal lesions involving keratinocytes. We examined here the signal transduction of lipopolysaccharide (LPS) in oral keratinocytes. Oral keratinocytes did not express CD14, but expression of CD58 and CD59 was observed by flow cytometry and reverse transcription-PCR. The binding between LPS and keratinocytes was strongly inhibited by pretreatment of keratinocytes with anti-CD59 monoclonal antibody (mAb) or phosphatidylinositol-specific phospholipase C (PI-PLC) but was not inhibited by anti-CD14 or anti-CD58 mAb. In LPS-treated keratinocytes, nuclear translocation of nuclear factor-kappa B (NF-kappaB) was induced and generation of granulocyte-macrophage colony-stimulating factor, interleukin-6 and tumour necrosis factor-alpha was enhanced. These upregulations in nuclear translocation of NF-kappaB and cytokine generation were not suppressed by anti-CD14 mAb or anti-CD58 mAb but were suppressed by anti-CD59 mAb and PI-PLC. Moreover, the transfection of CD59 antisense oligonucleotide into keratinocytes markedly suppressed LPS-induced nuclear translocation of NF-kappaB and cytokine generation. These results indicate that, through CD59, the LPS signal is transduced into the nucleus via NF-kappaB activation inducing cytokine generation, which may be involved in dermal and mucosal inflammatory diseases.
authors.affiliation: Department of Oral Surgery, Kochi Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan. yamamott@kochi-ms.ac.jp
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nakane
authors.forename: Sayoko
authors.initials: S
authors.lastname: Doi
authors.forename: Tokio
authors.initials: T
authors.lastname: Osaki
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D015152:Blotting, Northern; D018968:CD58 Antigens / Q000032:analysis / Q000235:genetics; D018957:CD59 Antigens / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000187:drug effects; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D004705:Endocytosis / Q000187:drug effects; D019286:Endopeptidase K / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods; D005786:Gene Expression Regulation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D015603:Keratinocytes / Q000737:chemistry* / Q000502:physiology; D018950:Lipopolysaccharide Receptors / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000166:cytology; D009055:Mouth / Q000166:cytology*; D016328:NF-kappa B / Q000032:analysis / Q000502:physiology; D043265:Phosphatidylinositol Diacylglycerol-Lyase / Q000494:pharmacology; D054801:Phosphoinositide Phospholipase C; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
chemical_list: D000906:Antibodies; D018968:CD58 Antigens; D018957:CD59 Antigens; D016373:DNA, Antisense; D015850:Interleukin-6; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D016328:NF-kappa B; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D054801:Phosphoinositide Phospholipase C; D019286:Endopeptidase K; D043265:Phosphatidylinositol Diacylglycerol-Lyase