Lee-Jung Chien
The Third Department of Medicine, School of Medicine,
Tokyo Medical and Dental University, Tokyo, Japan
Lee-Jung Chien
The Third Department of Medicine, School of Medicine,
Tokyo Medical and Dental University, Tokyo, Japan
| Objective: To investigate the adhesive interaction
of HL60 cells to vascular endothelium during their differentiation for
the purpose of establishment of a novel experimental model for E-selectin
dependent leukocyte-endothelial interaction.
Methods: HL60 cells were incubated in the presence of dimethylsulfoxide (DMSO) for 72hours to induce granulocytic differentiation. The adhesion assay of HL60 cells to cytokine activated (IL-1£], 10U/ml, 4hrs) human umbilical vein endothelial cells (HUVEC) were carried out under non-static (64rpm, 10min) and static assay conditions. Transmigration assay of HL60 cells were carried out using HUVEC plated on transwell culture inserts. Results: Granulocytic differentiation of HL60 cells significantly enhanced their adhesion to activated HUVEC. Preincubation of activated HUVEC with E-selectin mAb (H18/7) and incubation of differentiated HL60 (dHL60) and polymorphonuclear neutrophils (PMN) with a mAb against sialyl lewis X (sLx), a carbohydrate ligand of E-selectin, blocked adhesion to activated HUVEC. Transmigration of HL60 cells were significantly induced after differentiation. This induced transmigration of dHL60 but not that of PMN was blocked by preincubation of EC monolayer with H18/7. Conclusions: DMSO- induced differentiation of HL60 cells enhanced their adhesion to as well as transmigration across activated HUVEC via E-selectin dependent mechanism. This would be an important experimental model for E-selectin dependent leukocyte-EC interaction in inflammation. (J Intern Med Taiwan 1999;10:99-107)
|