CD40-CD40 ligand signal induces the intercellular adhesion molecule-1 expression through nuclear factor-kappa B p50 in cultured salivary gland epithelial cells from patients with Sjögren's syndrome.
The purpose of this study is to clarify signal transduction of expression of the intercellular adhesion molecule-1 (ICAM-1) via CD40-CD40 ligand in salivary glands of patients with Sjögren's syndrome (SS). We used cultured salivary gland epithelial cells (SG cells) from 15 SS patients and 8 controls obtained by labial minor salivary gland biopsy. First, ICAM-1 expression was determined with reverse transcriptase-polymerase chain reaction and flow cytometry in the presence or absence of soluble CD40L (sCD40L). Next, SG cells were transfected with plasmids of pGL1.3-Luc inserted with promoter region of ICAM-1, pGL1.3kB(-)-Luc mutated in nuclear factor kappa-B (NF-kappaB) binding site of pGL1.3-Luc and pNF-kappaB-Luc by lipofection method. Luciferase activity of the cells was measured in the presence or absence of sCD40L or sCD40L and an NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC). Finally NF-kappaB family proteins of cell nuclear extracts were determined. ICAM-1 expression was significantly enhanced with sCD40L at the mRNA and protein level. Activity of pNF-kappaB-Luc and pGL1.3-Luc was significantly elevated by stimulation with sCD40L and suppressed by PDTC. NF-kappaB p50 protein level was elevated by stimulation with sCD40L and suppressed by PDTC. Our results suggest that sCD40L enhances the ICAM-1 expression by activation of NF-kappaB p50 in the SS SG cells.
Saito
Miwako
M
Third Department of Internal Medicine, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu 431-3192, Japan. miwako.t@nifty.com
Ota
Yasuhiro
Y
Ohashi
Hiroyuki
H
Dei
Yoshiaki
Y
Shimoyama
Kumiko
K
Suzuki
Daisuke
D
Hayashi
Hideharu
H
Ogawa
Noriyoshi
N
D000328:Adult; D000368:Aged; D019013:CD40 Antigens / Q000276:immunology; D023201:CD40 Ligand / Q000502:physiology*; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D052002:NF-kappa B p50 Subunit / Q000502:physiology*; D012469:Salivary Glands / Q000166:cytology / Q000378:metabolism*; D015398:Signal Transduction; D012859:Sjogren's Syndrome / Q000276:immunology*; D015854:Up-Regulation
D019013:CD40 Antigens; D052002:NF-kappa B p50 Subunit; D018799:Intercellular Adhesion Molecule-1; D023201:CD40 Ligand