Glycosphingolipid composition of a new immortalized human cerebromicrovascular endothelial cell line

Previous studies have demonstrated the involvement of glycosphingolipid (GS L) antigens in the pathogenesis of immune-mediated neurological disorders s uch as peripheral neuropathies and multiple sclerosis. To study the role of the blood-brain barrier (BBB) in these disorders, we used a new human cere bromicrovascular endothelial cell (HCEC) line that has been immortalized th rough transfection with the plasmid pSV3-neo encoding for the SV40 large T- antigen and the neomycin gene. The immortalized HCEC (SV-HCEC) exhibited ac celerated proliferation rates but maintained phenotypic properties of early -passage control cells. Therefore, this human cell line may serve as a usef ul in vitro model for studying the properties of the human BBB. We first in vestigated 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 gly colipids), lactosylceramide (36%), globotriaosylceramide (3%), and globosid e (43%). Trace amounts of paragloboside, lactosaminyl paragloboside, and su lfoglucuronyl paragloboside could also be detected by TLC-immunostaining. T hese results provide the basis for further investigations of the expression of these cell surface antigens in cultured SV-HCECs on activation with inf lammatory cytokines such as interleukin-1 beta, tumor necrosis factor-alpha , and interferon-gamma, which have been implicated as playing an important role in the pathogenesis of many nervous system disorders.