A human T-cell receptor recognizes 'O'-linked sugars from the hinge region of human IgA1 and IgD.
A receptor which binds secretory IgA (sIgA) is expressed on human T cells from patients with systemic lupus erythematosus, rheumatoid arthritis, Behcet's syndrome and IgA nephropathy and on normal T cells following phytohaemagglutinin (PHA) stimulation. The specificity of this receptor was initially probed with a panel of normal serum immunoglobulins in competitive inhibition assays with sIgA using two-colour immunofluorescence. While the receptor showed the strongest affinity for IgA1 (IC(50)10(-6) M), IgD which has a similarly glycosylated hinge region to IgA1, also bound to the receptor (IC50 10(-5) M). IgA2, which lacks the 'O'-glycosylated hinge region, did not significantly inhibit the binding at these concentrations suggesting that the IgA determinants for this receptor might be the oligosaccharides present in the hinge region of IgA1. IgA1 has up to 10 'O'-linked oligosaccharides and four N-linked oligosaccharides per molecule. In order to probe the role of the 'O'-linked hinge sugars in the binding event, a sugar library was prepared from IgA1 by a procedure designed to release 'O'-linked oligosaccharides preferentially, and to retain them in the natural closed ring formation. The sugars were released by hydrazinolysis at 65 degrees and the resulting oligosaccharide library analysed by high voltage paper electrophoresis (HVE) and P4 gel permeation chromatography. Competitive inhibition studies demonstrated that both the library and the individual 'O'-linked sugars associated with IgA1 were implicated in the binding of IgA1 to this receptor (IC50 between 1 x 10(-5) M and 6 x 10(-5) M). Within this range the individual sugars showed small differences in their affinity for the receptor in the following order: Gal beta 3GalNAc = NeuNAc2 alpha 3(6)Gal beta 3GalNAc > NeuNAc2 alpha 3(6)Gal beta 3[NeuNAc2 alpha 6]GalNAc > or = GalNAc.
Rudd
P M
PM
Department of Biochemistry, University of Oxford, UK.
Fortune
F
F
Patel
T
T
Parekh
R B
RB
Dwek
R A
RA
Lehner
T
T
D001667:Binding, Competitive / Q000276:immunology; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D004590:Electrophoresis, Paper; D006801:Humans; D006834:Hydrazines; D007070:Immunoglobulin A / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D007071:Immunoglobulin A, Secretory / Q000378:metabolism; D007072:Immunoglobulin D / Q000378:metabolism*; D008213:Lymphocyte Activation / Q000276:immunology; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*
D006834:Hydrazines; D007070:Immunoglobulin A; D007071:Immunoglobulin A, Secretory; D007072:Immunoglobulin D; D009844:Oligosaccharides; D011134:Polysaccharides; D011948:Receptors, Antigen, T-Cell