Detection of gender difference and epitope specificity of IgG antibody activity against IgA1 hinge portion in IgA nephropathy patients by using synthetic hinge peptide and glycopeptide probes

Background and Aims: There are many reports on the presence of an incompletely glycosylated O-linked oligosaccharide(s) on the IgA1 hinge region in some immunoglobulin (IgA) nephropathy patients. Furthermore, the production of an antibody against the naked hinge peptide portion was reported in an IgA nephropathy patient. In this report, characterization of the IgG antibody against the hinge portion was carried out by using synthetic hinge glycopeptide probes. Methods and Results: The following synthetic hinge peptide and glycopeptides were prepared: 19mer peptide, V-P-S-T-P-P-T-P-S-P-S-T-P-P-T-P-S-P-S (designated HP), the peptide having a single alpha-linked GalNAc residue at positions 4, 7, 9, 11 and 15 (4 GN - 15 GN, respectively) and the same peptide having five GalNAc residues at all five positions (GN5). The mean value of the antibody activity against these probes was compared with each other. The highest activity against the naked hinge peptide (HP) and lowest activity against the fully glycosylated hinge peptide (GN5) were obvious. As attachment of GalNAc to position 4 or 11 on the peptide brought about a significant reduction of the activity against the naked hinge peptide, the P-S-T-P sequence included in both positions was thought to be the most probable site recognized by these antibodies. As an additional unexpected observation, a gender difference in this antibody activity against all the probes was found. The antibody activity in a female was significantly higher compared with that in a male. Conclusion: Because the frequency of incidence of IgA nephropathy is known to be slightly higher in males, this gender difference might indicate a protective meaning to remove aberrantly glycosylated molecules from the patient's serum.