Glycosphingolipid composition of a new immortalized human cerebromicrovascular endothelial cell line.
Previous studies have demonstrated the involvement of glycosphingolipid (GSL) antigens in the pathogenesis of immune-mediated neurological disorders such as peripheral neuropathies and multiple sclerosis. To study the role of the blood-brain barrier (BBB) in these disorders, we used a new human cerebromicrovascular endothelial cell (HCEC) line that has been immortalized through transfection with the plasmid pSV3-neo encoding for the SV40 large T-antigen and the neomycin gene. The immortalized HCEC (SV-HCEC) exhibited accelerated proliferation rates but maintained phenotypic properties of early-passage control cells. Therefore, this human cell line may serve as a useful in vitro model for studying the properties of the human BBB. We first investigated the GSL composition of cultured SV-HCECs. The major gangliosides were GM3 (62% of total gangliosides), GM2 (18%), GM1 (3%), and GD1a (15%). The major neutral GSLs were glucosylceramide (15% of the total neutral glycolipids), lactosylceramide (36%), globotriaosylceramide (3%), and globoside (43%). Trace amounts of paragloboside, lactosaminyl paragloboside, and sulfoglucuronyl paragloboside could also be detected by TLC-immunostaining. These results provide the basis for further investigations of the expression of these cell surface antigens in cultured SV-HCECs on activation with inflammatory cytokines such as interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma, which have been implicated as playing an important role in the pathogenesis of many nervous system disorders.
Duvar
S
S
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia, USA.
Suzuki
M
M
Muruganandam
A
A
Yu
R K
RK
D001812:Blood-Brain Barrier; D002460:Cell Line; D002540:Cerebral Cortex / Q000098:blood supply* / Q000166:cytology; D002855:Chromatography, Thin Layer; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000166:cytology*; D005732:Gangliosides / Q000032:analysis; D005915:Globosides / Q000032:analysis; D006028:Glycosphingolipids / Q000032:analysis*; D006801:Humans; D008833:Microcirculation / Q000166:cytology; D016339:Spectrometry, Mass, Fast Atom Bombardment
D005732:Gangliosides; D005915:Globosides; D006028:Glycosphingolipids; C051917:sulfate-3-glucuronyl paragloboside