title: The effect on IgG glycosylation of altering beta1, 4-galactosyltransferase-1 activity in B cells.
abstract: An absence of galactose on the N-linked oligosaccharides of immunoglobulin G (IgG) has been shown to affect the functional activity of the antibody molecule. In patients with rheumatoid arthritis there is an increased proportion of IgG which lacks galactose and correspondingly lower levels of beta1, 4-galactosyltransferase (beta4Gal-T) activity. The recent demonstration of several expressed beta4Gal-T genes in man raises the possibility that the enzyme responsible for the decreased IgG galactose is not the "classical" beta4Gal-T (beta4Gal-T1). To directly address the question of whether reduced beta4Gal-T1 would lead to reduced IgG galactose, the level of beta4Gal-T1 in a human IgG-secreting B cell line was specifically altered using stable transfection with sense (SpcDNA3-Gal-T1) or antisense (ASpcDNA3-Gal-T1) human beta4Gal-T1 cDNA. SpcDNA3-Gal-T1 B cell transfectants expressed up to a 2.5-fold higher level of beta4Gal-T enzyme activity for the exogenous neoglycoconjugate acceptor GlcNAc-pITC-BSA than did ASpcDNA3-Gal-T1 transfectants. Flow cytometric analysis with Ricinus communis agglutinin I (RCAI) revealed an overall greater number of Galbeta1,4GlcNAc structures in the fixed and permeabilized SpcDNA3-Gal-T1 B cell transfectants compared with the ASpcDNA3-Gal-T1 transfectants. Moreover, there was increased galactosylation of IgG secreted from the SpcDNA3-Gal-T1 transfectants relative to the ASpcDNA3-Gal-T1 B cell transfectants. Alteration of the level of the "classical" beta4Gal-T (beta4Gal-T1) in B cells therefore affects IgG glycosylation.
authors.affiliation: Department of Immunology, University College London, Windeyer Building, 46 Cleveland Street, London W1P 6DB, UK.
authors.forename: J
authors.initials: J
authors.lastname: Keusch
authors.forename: P M
authors.initials: PM
authors.lastname: Lydyard
authors.forename: P J
authors.initials: PJ
authors.lastname: Delves
mesh_terms: D001402:B-Lymphocytes / Q000201:enzymology*; D005434:Flow Cytometry; D005690:Galactose / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D009237:N-Acetyllactosamine Synthase / Q000235:genetics* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014162:Transfection / Q000235:genetics
chemical_list: D006023:Glycoproteins; D007074:Immunoglobulin G; D016376:Oligonucleotides, Antisense; D012710:Serum Albumin, Bovine; D009237:N-Acetyllactosamine Synthase; D005690:Galactose