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 language | English (US) |
|---|---|
| Pages (from-to) | 1970-1976 |
| Number of pages | 7 |
| Journal | Journal of Neurochemistry |
| Volume | 75 |
| Issue number | 5 |
| DOIs | |
| State | Published - Jan 1 2000 |
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Keywords
- Antibodies
- Blood-brain barrier
- Endothelial cells
- Fast-atom bombardment mass spectrometry
- Glycosphingolipids
ASJC Scopus subject areas
- Biochemistry
- Cellular and Molecular Neuroscience
Cite this
Glycosphingolipid composition of a new immortalized human cerebromicrovascular endothelial cell line. / Duvar, S.; Suzuki, M.; Muruganandam, A.; Yu, Robert K.
In: Journal of Neurochemistry, Vol. 75, No. 5, 01.01.2000, p. 1970-1976.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Glycosphingolipid composition of a new immortalized human cerebromicrovascular endothelial cell line
AU - Duvar, S.
AU - Suzuki, M.
AU - Muruganandam, A.
AU - Yu, Robert K
PY - 2000/1/1
Y1 - 2000/1/1
N2 - 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.
AB - 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.
KW - Antibodies
KW - Blood-brain barrier
KW - Endothelial cells
KW - Fast-atom bombardment mass spectrometry
KW - Glycosphingolipids
UR - http://www.scopus.com/inward/record.url?scp=0033793238&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033793238&partnerID=8YFLogxK
U2 - 10.1046/j.1471-4159.2000.0751970.x
DO - 10.1046/j.1471-4159.2000.0751970.x
M3 - Article
C2 - 11032886
AN - SCOPUS:0033793238
VL - 75
SP - 1970
EP - 1976
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
SN - 0022-3042
IS - 5
ER -