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Characterization of...
Abstract
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- The alpha1,3-fucosyltransferase VII (Fuc-TVII) is involved in the biosynthesis of E- and P-selectin ligands such as sialyl Lewis x (SLe(x)) on human leukocytes. Recently, individuals were characterized carrying a missense mutation (G329A; Arg110-Gln) in the FUT7 gene encoding this enzyme. The mutated FUT7 construct produced a Fuc-TVII enzyme with impaired activity compared with the wildtype enzyme. Polymorphonuclear granulocytes from an individual carrying this mutation homozygously also showed a reduced expression of SLe(x). In the present study, we have established Epstein-Barr virus-transformed B-cell lines from this individual (SIGN) and from an individual not carrying the mutation (IWO). The cell lines were confirmed to be of B-cell origin by flow cytometry analysis. IWO cells interacted with E-selectin in an in vitro flow chamber analysis whereas SIGN cell did not. However, when SIGN cell was transiently transfected with wildtype FUT7 cDNA, interaction with E-selectin could be restored. Cell surface expression of the SLe(x)-related epitopes recognized by antibodies CSLEX-1, KM-93 and HECA-452 was elevated on IWO cells compared with that on SIGN cells, consistent with a role of these antigens in E-selectin recognition. These cell lines will be useful in further characterization of E-selectin ligands and encourage further studies on the consequences of the FUT7-G329A mutation in vivo.
Keyword
- B-Lymphocytes/drug effects/*enzymology/immunology
- Carbohydrates/analysis
- *Cell Line
- Transformed
- DNA
- Complementary/genetics
- E-Selectin/*biosynthesis/pharmacology
- Epitopes
- B-Lymphocyte/analysis
- Fucosyltransferases/*genetics
- Gene Expression
- Herpesvirus 4
- Human/*physiology
- Homozygote
- Humans
- Ligands
- Mutation
- Missense
- Oligosaccharides/analysis
- RNA
- Messenger/analysis/metabolism
- Transfection
- MEDICINE
Publication and Content Type
- ref (subject category)
- art (subject category)
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