Activation of human endothelial cells by tumor necrosis factor-alpha results in profound changes in the expression of glycosylation-related genes.
The endothelium plays a central role in the logistics of the immune system by allowing the selective transmigration of leukocytes, as well as the maintenance of the circulation and coagulation homeostasis. Evidence is increasing that the carbohydrate composition of the endothelial cell surface is critical for the cells to exert their physiological function. The major aim of this study is to unravel the mechanisms underlying the expression of carbohydrate structures by endothelial cells, which are involved in leukocyte adhesion and migration. Using quantitative real-time PCR, the expression profile of a selected group of 74 glycosylation-related genes has been determined in human umbilical vein endothelial cells (HUVEC) and human foreskin microvascular endothelial cells (FMVEC) under control and TNFalpha-induced conditions. The set of genes comprised 59 glycosyltransferases, 6 mannosidases and 9 sulfotransferases. In parallel, the overall cell surface glycan profile has been assessed by the use of glycan-specific lectins and monoclonal antibodies. The results demonstrate that HUVEC and FMVEC differ substantially in the expression of glycosylation-related genes and, accordingly, also in the presence of different glycan epitopes on the cell membrane. Induction of an inflamed phenotype of the cells by treatment with TNFalpha differentially modulates a set of these genes in HUVEC and FMVEC resulting in a change in the cell membrane associated glycans that are of importance in inflammation-related endothelial cell-surface processes.
García-Vallejo
Juan J
JJ
Department of Molecular Cell Biology & Immunology, VU Medical Center, Amsterdam, The Netherlands. jj.garciavallejo@vumc.nl
Van Dijk
Willem
W
Van Het Hof
Bert
B
Van Die
Irma
I
Engelse
Marten A
MA
Van Hinsbergh
Victor W M
VW
Gringhuis
Sonja I
SI
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000276:immunology* / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology; D000939:Epitopes; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006031:Glycosylation; D006801:Humans; D007962:Leukocytes / Q000166:cytology / Q000276:immunology; D008969:Molecular Sequence Data; D011485:Protein Binding; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D000939:Epitopes; D014409:Tumor Necrosis Factor-alpha