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Characterization of EBV-transformed B-cells established from an individual homozygously mutated (G329A) in the FUT7 alpha1,3-fucosyltransferase gene

Journal article
Authors P. Bengtson
Henrik Zetterberg
T. Mellberg
P. Pahlsson
Göran Larson
Published in Scand J Immunol
Volume 62
Issue 3
Pages 251-8
Publication year 2005
Published at Institute of Laboratory Medicine, Dept of Clinical Chemistry/Transfusion Medicine
Pages 251-8
Language en
Links dx.doi.org/10.1111/j.1365-3083.2005...
Keywords 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
Subject categories Medical and Health Sciences

Abstract

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.

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