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Glycosphingolipid Composition of a New Immortalized Human Cerebromicrovascular Endothelial Cell Line
Author(s) -
Duvar Sevim,
Suzuki Minoru,
Muruganandam Arumugam,
Yu Robert K.
Publication year - 2000
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.2000.0751970.x
Subject(s) - glycosphingolipid , globotriaosylceramide , lactosylceramide , immortalised cell line , glycolipid , biology , cell culture , immunostaining , antigen , microbiology and biotechnology , immunology , cell , biochemistry , medicine , pathology , fabry disease , genetics , immunohistochemistry , disease
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‐1β, tumor necrosis factor‐α, and interferon‐γ, which have been implicated as playing an important role in the pathogenesis of many nervous system disorders.