2001 Fiscal Year Final Research Report Summary
Role of leukocyte-endothelial interaction for pathogenesis of vessel diseases perturbed by cytokines
| Project/Area Number |
12470530
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Laboratory medicine
|
|---|
| Research Institution | Keio University |
|---|
Principal Investigator |
WATANABE Kiyoaki Keio University, School of Medicine, Professor, 医学部, 教授 (20101983)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KAWAI Yohko Keio University, School of Medicine, Assistant Professor, 医学部, 講師 (00129727)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Keywords | endothelium / leukocyte / cytokine / tumor necrosis factor (TNF) / NF-κB / CD146 (Mgl-CAM) / selectin family / immunoglobulin gene super-family |
|---|
| Research Abstract |
To investigate the role of leukocyte-endothelial interaction for pathogenesis of vessel diseases perturbed by cytokines, in vitro experiments have been undertaken. Hemodynamic forces modulate various endothelial cell functions even in the presence of cytokines under gene regulation. We have investigated the effect of shear stress on coagulation and fibrinolysis system in cultured human umbilical vein endothelial cells (HUVECs) perturbed by cytokines or lipopolysaccharide (LPS), using modified cone-plate type viscometer, in which well controlled and defined shear forces were generated. Several monoclonal antibodies have been developed against human nasal-mucous microvessels. Monoclonal antibodies have been produced against two adhesive glycoproteins for Eselectin and Mel-CAM that act as important molecules for leucocyte adhesion to endothelium under inflammatory site. E-selectin was not detected in resting unstimulated HUVECs, whereas, surface membrane expression of E-selectin was incre
… More
ased 3〜6 hours after the stimulation of TNF. In contrast, Mel-CAM expression was clearly observed in resting unstimulated HUVECs, whereas the release to culture supernatant was increased after TNF stimulation and decreased under shear stress. We also investigated the expression of ICAM-1 and VCAM-1 in HUVECs, using western blotting. ICAM-1 and VCAM-1 was not detected in resting unstimulated HUVECs, whereas, the expression of ICAM-1 and VCAM-1 have been increased 3〜6 hours after the stimulation of TNF. We have isolated mononuclear cells (MNCs) from human peripheral blood, using Ficoll-gradient methods. No adhesion of MNCs was observed to resting unstimulated HUVECs, whereas, the leukocyte adhesion to HUVECs was clearly observed 6〜9 hours after TNF-stimulation. Interestingly the expression of these cytoadhesion molecules and leukocyte adhesion to TNF-stimulated HUVECs have been decreased by the inhibition of NF-κB activation. These results indicate that the regulation of leukocyte-endothelial interaction under TNF-stimulation through NF-κB activation might be very important modulator for inflammation and atheroclerosis through cytoadhesion molecules. Less
|
