Tissue factor expression in vascular endothelial cells induced by allogeneic T cells
10.3969/j.issn.1673-8225.2009.53.017
- VernacularTitle:异基因T淋巴细胞诱导血管内皮细胞组织因子的表达
- Author:
Jian ZHANG
;
Quan LI
;
Weiming LI
;
Ping ZOU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(53):10481-10486
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Host endothelial cells are important targets of alloreactive donor cytotoxic T lymphocytes in graft-versus-host disease (GVHD). And the expression of tissue factor in vascular endothelial cells is up-regulated post-injury and activation. So tissue factor expression in vascular endothelial cells may play a pivotal role in the pathogenesis of GVHD.OBJECTIVE: To investigate the molecular mechanism of allogeneic T cell-induced tissue factor (TF) expression in vascular endothelial cells.DESIGN, TIME AND SETTING: The cytology observation was performed at immunological laboratory of ongji Medical College, Huazhong University of Science and Technology from March to October 2008.MATERIALS: Human primary umbilical vein endothelial cells (HUVECs) were purchased from Science cell laboratory (CA, USA).METHODS: Allogeneic CD4~+ and CD8~+T cells were isolated from peripheral blood mononuclear cells (PBMC) by positive selection using magnetic beads coated with anti-CD4 or anti-CD8 antibody. After thorough washing with PBS, 1×10~6 allogeneic CD4~+T cells or CD8~+T cells were added to HUVECs (ratio 10:1). The cells were grown at 37 ℃ in a humidified atmosphere containing c-jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated protein kinase (ERK). HUVECs were pretreated with TF antibody 1 hour before co-culturing; the untreated HUVECs were used as controls. MAIN OUTCOME MEASURES: Allogeneic T cells-induced TF expression in vascular endothelial cells was determined by real-time PCR and flow cytometry. Western blot was used to examine allogeneic T cells-induced phosphorylation of MAPK in HUVEC.RESULTS: After co-culturing with allogeneic CD4~+T cells or CD8~+T cells for 6, 12, and 24 hours, the expression rates of TF on HUVEC membrane were obvious increased than that of the control group (P < 0.05), the expression levels of TF mRNA in HUVEC were also significantly elevated (P < 0.05). Allogeneic CD4~+T cells-induced expression rates of TF were significantly higher than allogeneic CD8~+T cells-induced expression rates at 12 and 24 hours after culture. The expression of p38MAPK and JNK was enhanced after culture with allogeneic T cells, yet the ERK expression had no changes. The p38MAPK or JNK can notably down regulated TF expression, which could block the TF expression when worked together. CONCLUSION: Allogeneic T cells induced TF expression in HUVECs via activation of p38MAPK and JNK.
