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Characterization of EBV‐transformed B‐cells Established from an Individual Homozygously Mutated (G329A) in the FUT7 α1,3‐fucosyltransferase Gene
Author(s) -
Bengtson P.,
Zetterberg H.,
Mellberg T.,
Påhlsson P.,
Larson G.
Publication year - 2005
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.2005.01650.x
Subject(s) - fucosyltransferase , biology , microbiology and biotechnology , mutation , missense mutation , cell culture , transfection , gene , antibody , epitope , genetics
The α1,3‐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; Arg 110 ‐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 .