Complement activation in experimental IgA nephropathy: An antigen-mediated process

Abstract

Complement activation associated with immune complex glomerular deposition plays an important role in renal injury. In the present studies we performed three series of experiments to identify how IgA immune complexes activate complement. The first series of experiments was designed to determine whether the presence of an antigen within a glomerular IgA immune deposit is required for complement activation. In these experiments, large-sized covalently cross-linked IgA oligomers (X-IgA) were prepared with purified IgA anti-dinitrophenyl (DNP) and a bivalent affinity-labeling antigen, bis-2,4-DNP-pimelic acid ester. These X-IgA oligomers have free antigen-binding sites that will bind DNP-conjugated antigens. Two groups of mice were treated with either X-IgA or X-IgA followed, after two hours, by an antigen DNP-Ficoll. Immunofluorescent examination of renal tissues, obtained six hours after the initial injection, revealed an equal intensity of IgA glomerular deposits in both groups of mice. Glomerular C3 deposits were only detectable in the renal tissues of mice that had DNP-Ficoll bound to X-IgA. In the second series of experiments, a pair of preformed IgA immune complexes, differing only in one antigenic structural feature (DNP), were used to examine the role of the antigen in inducing glomerular C3 deposits in two groups of mice. These pre-formed immune complexes were prepared with IgA anti-phosphorylcholine (PC) and either PC-conjugated to bovine serum albumin (PC-BSA) or PC-BSA which was further modified with DNP (PC/DNP-BSA). Although the IgA immunofluorescent intensity and pattern in the glomerular deposits were equivalent for both groups, intense C3 deposits were exclusively associated with the PC/DNP-BSA-containing immune complexes. Analysis of the relative conversion of normal human serum C3 to inactive C3b (iC3b) by X-IgA, various antigens and their respective IgA immune complexes was highly dependent on the nature of the antigen.(ABSTRACT TRUNCATED AT 250 WORDS)