Glycosphingolipid composition of a new immortalized human cerebromicrovascular endothelial cell line

S. Duvar, M. Suzuki, A. Muruganandam, R. K. Yu

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1970-1976
Number of pages7
JournalJournal of Neurochemistry
Volume75
Issue number5
DOIs
StatePublished - 2000

Keywords

  • Antibodies
  • Blood-brain barrier
  • Endothelial cells
  • Fast-atom bombardment mass spectrometry
  • Glycosphingolipids

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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