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Effects of TNF- on Leukocyte Adhesion Molecule Expressions in Cultured Human Lymphatic Endothelium

Yoshihiko Sawa, Yukitaka Sugimoto, Takeshi Ueki, Hiroyuki Ishikawa, Atuko Sato, Toshikazu Nagato and Shigemitsu Yoshida

Department of Morphological Biology (YS,YS,AS,TN) and Department of Oral Growth & Development (TU,HI), Fukuoka Dental College, Fukuoka, Japan, and Department of Oral Functional Science, Graduate School of Dental Medicine, Hokkaido University, Sapporo, Japan (SY)

Correspondence to: Yoshihiko Sawa, Department of Morphological Biology, Fukuoka Dental College, 2-15-1 Tamura, Sawara-Ku, Fukuoka 814-0193, Japan. E-mail: ysawa{at}college.fdcnet.ac.jp

TNF- alters leukocyte adhesion molecule expression of cultured endothelial cells like human umbilical vein endothelial cells (HUVEC). This study was designed to investigate the changes in vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and platelet endothelial cell adhesion molecule-1 (PECAM-1) expression with TNF- stimulation in cultured human neonatal dermal lymphatic endothelial cells (HNDLEC). The real-time quantitative PCR analysis on HNDLEC showed that TNF- treatment leads to increases of VCAM-1 and ICAM-1 mRNAs to the 10.8- and 48.2-fold levels of untreated cells and leads to a reduction of PECAM-1 mRNA to the 0.42-fold level of untreated cells. Western blot and immunohistochemical analysis showed that TNF- leads to VCAM-1 and ICAM-1 expressions that were inhibited by antiserum to human TNF receptor or by AP-1 inhibitor nobiletin. In flow cytometry analysis, the number of VCAM-1- and ICAM-1-positive cells increased, and PECAM-1-positive cells decreased with TNF- treatment. Regarding protein amounts produced in cells and amounts expressed on the cell surface, VCAM-1 and ICAM-1 increased in HNDLEC and HUVEC, and PECAM-1 decreased in HNDLEC in a TNF- concentration-dependent manner. VCAM-1, ICAM-1, and PECAM-1 protein amounts in TNF--stimulated cells were lower in HNDLEC than in HUVEC. This suggests that the lymphatic endothelium has the TNF--induced signaling pathway, resulting in increased VCAM-1 and ICAM-1 expression to a weaker extent than blood endothelium and PECAM-1 reduction to a stronger extent than blood endothelium. (J Histochem Cytochem 55:721–733, 2007)

Key Words: lymphatic endothelium • TNF- • platelet endothelial cell adhesion molecule-1 • intercellular adhesion molecule-1 • vascular cell adhesion molecule-1

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