Characterization of EBV-transformed B-cells established from an individual homozygously mutated (G329A) in the FUT7 alpha1,3-fucosyltransferase gene.
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.
Bengtson
P
P
Department of Biomedicine and Surgery, Division of Cell Biology, Linköping University, Linköping;, Sweden.
Zetterberg
H
H
Mellberg
T
T
Påhlsson
P
P
Larson
G
G
D001402:B-Lymphocytes / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D002241:Carbohydrates / Q000032:analysis; D002461:Cell Line, Transformed; D018076:DNA, Complementary / Q000235:genetics; D019040:E-Selectin / Q000096:biosynthesis* / Q000494:pharmacology; D018985:Epitopes, B-Lymphocyte / Q000032:analysis; D005647:Fucosyltransferases / Q000235:genetics*; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006720:Homozygote; D006801:Humans; D008024:Ligands; D020125:Mutation, Missense; D009844:Oligosaccharides / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen; D014162:Transfection
D002241:Carbohydrates; D018076:DNA, Complementary; D019040:E-Selectin; D018985:Epitopes, B-Lymphocyte; D008024:Ligands; D009844:Oligosaccharides; D012333:RNA, Messenger; D000080506:Sialyl Lewis X Antigen; C502597:FUT7 protein, human; D005647:Fucosyltransferases