Characterization of Anti-Mesangial Cell Antibodies and Their Target Antigens in Patients with Lupus Nephritis

Abstract

The pathogenetic mechanisms of lupus nephritis (LN) remain to be elucidated. In our previous study, autoantibodies against human glomerular mesangial cells (HMC) were identified in sera of most patients with lupus nephritis. The current study is to investigate the binding characteristics of anti-mesangial cell antibodies to human mesangial cell membrane. Serum samples were collected from 54 patients with renal biopsy proven lupus nephritis, 12 patients with systemic lupus erythematosus without clinical renal involvement, and 15 healthy subjects. Membrane proteins were obtained from in vitro cultured HMC by sonication and sequential centrifugation. DNase I were employed to remove DNA fragments in sera and membrane protein preperation and IgG F(ab')2 was obtained by pepsin digestion. Western Blot analysis was used to characterize the antibody and antigen interaction. In results, 25 of 54 (46.3%) sera from patients with lupus nephritis had anti-mesangial cell antibodies targeted at 74 kDa, 63 kDa, 52 kDa and 42 kDa protein bands of HMC membrane. Only four of 12 (33.3%) sera from patients without renal involovement recognized the protein bands at 74 kDa and 63 kDa, but not 52 kDa and 42 kDa. DNase treatment of the HMC membrane and the sera did not affect the binding. IgG F(ab')2 from sera of 10 patients with positive anti-mesangial cell antibodies could still bind the 63 kDa protein. In conclusion, anti-mesangial cell antibodies from sera of patients with lupus nephritis could bind membrane proteins of HMC directly without a DNA bridge and the binding was through antigen-antibody interation. Anti-mesangial cell antibodies might play some role in the pathogenesis of lupus nephritis(LN).