Effects of chemoattractant function of regulated upon activation normal T cell expressed and secreted with adenovirus-mediated gene transfer of S24F of the human umbilical vein endothelial cells

Abstract

Objective To investigate the effects of chemoattractant function of regulated upon activation normal T cell expressed and secreted (RANTES) with adenovirus-mediated gene transfer of S24F of the human umbilical vein endothelial cells (HUVECs).Methods Construction of the recombinant Adenoviruses Vector pAV-MCMV/S24F-GFP-3FLAG (Ad-S24F),an adenoviral vector containing no transgene pAV-MCMV-GFP (Ad-Null) was used as a control.HUVECs were isolated and cultured in vitro,and were transfected with Ad-S24F (Ad-S24F group),Ad-Null (Ad-Null group),or transfected with no virus (control group).The expression of reconstructive protein after transfection was detected by using Western blotting and fluorescence microscopy in each group.The in vitro transendothelialchemotaxis assay was used to compare RANTES-induced transmigration of peripheral blood mononuclear cells (PBMCs) across HUVECs cultured on the upper Transwell chamber.Results The successful construction of recombinant adenoviral vector carrying S24F gene.RANTES-induced PBMC transendothelialchemotaxis is inhibited by S24F [Ad-S24F:(9.20 ± 0.02) ％ ; Ad-Null:(17.70 ± 0.02) ％ ; control:(15.10 ± 0.01) ％].Conclusion Adenovirus-mediated gene transfer of S24F of the human umbilical vein endothelial cells may inhibit RANTES-induced PBMC transendothelialchemotaxis. Key words: S24F gene; Regulated upon activation normal T cell expressed and secreted; Adenovirus vector; Gene transfer