Toll-like receptor in salivary glands from patients with Sjögren's syndrome: functional analysis by human salivary gland cell line

J Rheumatol. 2007 May;34(5):1019-26. Epub 2007 Apr 1.

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.

Conclusion: 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.

MeSH terms

  • Adult
  • Aged
  • Apoptosis / drug effects
  • Cell Line
  • Dose-Response Relationship, Drug
  • Drug Combinations
  • Female
  • Humans
  • Immunoglobulin M / pharmacology
  • Intercellular Adhesion Molecule-1 / metabolism
  • Interleukin-6 / metabolism
  • Ligands
  • Lipopolysaccharides / pharmacology
  • Male
  • Middle Aged
  • Mitogen-Activated Protein Kinases / metabolism
  • Myeloid Differentiation Factor 88 / metabolism
  • Peptidoglycan / pharmacology
  • Poly I-C / pharmacology
  • Salivary Glands, Minor / drug effects
  • Salivary Glands, Minor / metabolism*
  • Salivary Glands, Minor / pathology
  • Sjogren's Syndrome / metabolism*
  • Sjogren's Syndrome / pathology
  • TNF-Related Apoptosis-Inducing Ligand / pharmacology
  • Toll-Like Receptors / metabolism*
  • fas Receptor / immunology

Substances

  • Drug Combinations
  • IL6 protein, human
  • Immunoglobulin M
  • Interleukin-6
  • Ligands
  • Lipopolysaccharides
  • MYD88 protein, human
  • Myeloid Differentiation Factor 88
  • Peptidoglycan
  • TNF-Related Apoptosis-Inducing Ligand
  • TNFSF10 protein, human
  • Toll-Like Receptors
  • fas Receptor
  • Intercellular Adhesion Molecule-1
  • Mitogen-Activated Protein Kinases
  • Poly I-C