title: Toll-like receptor in salivary glands from patients with Sjögren's syndrome: functional analysis by human salivary gland cell line.
abstract: OBJECTIVE
  We investigated expression of toll-like receptor (TLR) in labial salivary glands of patients with Sjögren's syndrome (SS) and functional TLR expression in the cultured salivary gland cell line.
  
  
  METHODS
  Expression of TLR2, TLR3, TLR4, and myeloid differentiation factor 88 (MyD88) in labial salivary glands was examined by immunohistochemistry. Human salivary gland (HSG) cell-line cells were cultured with TLR ligands [peptidoglycan, poly (I:C) and lipopolysaccharide], and CD54 expression and interleukin 6 (IL-6) production was studied. Phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, and Akt was examined by Western blotting. Activation of nuclear factor-kappaB (NF-kappaB) p65 in HSG cells was studied by NF-kappaB p65 nuclear translocation by microscopic immunofluorescence or chemiluminescent electrophoretic mobility shift assay and detection of NF-kappaB p65 phosphorylation.
  
  
  RESULTS
  TLR2, TLR3, TLR4, and MyD88 were more strongly expressed in the labial salivary glands of SS patients (n =12) than in control subjects (n = 4), and were found in salivary-infiltrating mononuclear cells as well as acinar cells and ductal epithelial cells. In cultured HSG cells, a similar expression pattern was observed, and TLR ligands stimulated CD54 expression and IL-6 production. TLR ligands induced phosphorylation of ERK, JNK, and p38 in HSG cells, but not Akt phosphorylation or activation of NF-kappaB p65.
  
  
  CONCLUSIONS
  Although the putative ligands remain to be determined, our study indicated the activation of the TLR-mediated immune response in SS, and suggested that the TLR effect is mediated through the mitogen-activated protein kinase pathway.
authors.affiliation: First Department of Internal Medicine, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. atsushik@net.nagasaki-u.ac.jp
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kawakami
authors.forename: Koto
authors.initials: K
authors.lastname: Nakashima
authors.forename: Mami
authors.initials: M
authors.lastname: Tamai
authors.forename: Hideki
authors.initials: H
authors.lastname: Nakamura
authors.forename: Nozomi
authors.initials: N
authors.lastname: Iwanaga
authors.forename: Keita
authors.initials: K
authors.lastname: Fujikawa
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Aramaki
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Arima
authors.forename: Naoki
authors.initials: N
authors.lastname: Iwamoto
authors.forename: Kunihiro
authors.initials: K
authors.lastname: Ichinose
authors.forename: Makoto
authors.initials: M
authors.lastname: Kamachi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ida
authors.forename: Tomoki
authors.initials: T
authors.lastname: Origuchi
authors.forename: Katsumi
authors.initials: K
authors.lastname: Eguchi
mesh_terms: D000328:Adult; D000368:Aged; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D005260:Female; D006801:Humans; D007075:Immunoglobulin M / Q000494:pharmacology; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008024:Ligands; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D010457:Peptidoglycan / Q000494:pharmacology; D011070:Poly I-C / Q000494:pharmacology; D012470:Salivary Glands, Minor / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D012859:Sjogren's Syndrome / Q000378:metabolism* / Q000473:pathology; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000494:pharmacology; D051193:Toll-Like Receptors / Q000378:metabolism*; D019014:fas Receptor / Q000276:immunology
chemical_list: D004338:Drug Combinations; C508600:IL6 protein, human; D007075:Immunoglobulin M; D015850:Interleukin-6; D008024:Ligands; D008070:Lipopolysaccharides; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D010457:Peptidoglycan; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D051193:Toll-Like Receptors; D019014:fas Receptor; D018799:Intercellular Adhesion Molecule-1; D020928:Mitogen-Activated Protein Kinases; D011070:Poly I-C