IgA nephropathy and Henoch-Schoenlein purpura nephritis: aberrant glycosylation of IgA1, formation of IgA1-containing immune complexes, and activation of mesangial cells.
IgA1 in the circulation and glomerular deposits of patients with IgA nephropathy (IgAN) is aberrantly glycosylated; the hinge-region O-linked glycans are galactose-deficient. The circulating IgA1 of patients with Henoch-Schoenlein purpura nephritis (HSPN) has a similar defect. This aberrancy exposes N-acetylgalactosamine-containing neoepitopes recognized by naturally occurring IgG or IgA1 antibodies resulting in formation of immune complexes. IgA1 contains up to six O-glycosylation sites per heavy chain; it is not known whether the glycosylation defect occurs randomly or preferentially at specific sites. We sought to define the aberrant glycosylation of a galactose-deficient IgA1 myeloma protein and analyze the formation of the immune complexes and their biological activities. Supplementation of serum or cord-blood serum with this IgA1 protein resulted in formation of new IgA1 complexes. These complexes stimulated proliferation of cultured human mesangial cells, as did the naturally-occurring IgA1-containing complexes from sera of patients with IgAN and HSPN. Uncomplexed IgA1 did not affect cellular proliferation. Using specific proteases, lectin Western blots, and mass spectrometry, we determined the O-glycosylation sites in the hinge region of the IgA1 myeloma protein and IgA1 proteins from sera of IgAN patients. The IgA1 myeloma protein had galactose-deficient sites at residues 228 and/or 230 and 232. These sites reacted with IgG specific to galactose-deficient IgA1. IgA1 from the IgAN patients had galactose-deficient O-glycans at the same residues. In summary, we identified the neoepitopes on IgA1 responsible for formation of the pathogenic immune complexes. These studies may lead to development of noninvasive diagnostic assays and future disease-specific therapy.
Novak
Jan
J
University of Alabama at Birmingham, Department of Microbiology, Birmingham, AL 35294, USA. jannovak@uab.edu
Moldoveanu
Zina
Z
Renfrow
Matthew B
MB
Yanagihara
Takeshi
T
Suzuki
Hitoshi
H
Raska
Milan
M
Hall
Stacy
S
Brown
Rhubell
R
Huang
Wen-Qiang
WQ
Goepfert
Alice
A
Kilian
Mogens
M
Poulsen
Knud
K
Tomana
Milan
M
Wyatt
Robert J
RJ
Julian
Bruce A
BA
Mestecky
Jiri
J
D000936:Antigen-Antibody Complex / Q000276:immunology*; D005920:Glomerular Mesangium / Q000276:immunology / Q000378:metabolism; D005922:Glomerulonephritis, IGA / Q000276:immunology*; D006031:Glycosylation; D006801:Humans; D011695:IgA Vasculitis / Q000150:complications / Q000276:immunology*; D007070:Immunoglobulin A / Q000276:immunology / Q000378:metabolism*; D009393:Nephritis / Q000209:etiology / Q000276:immunology*
D000936:Antigen-Antibody Complex; D007070:Immunoglobulin A