Isotypes of autoantibodies against novel differential 4-hydroxy-2-nonenal-modified peptide adducts in serum is associated with rheumatoid arthritis in Taiwanese women

Rheumatoid arthritis (RA) is an autoimmune disorder with systemic inflammation and may be induced by oxidative stress that affects an inflamed joint. Our objectives were to examine isotypes of autoantibodies against 4-hydroxy-2-nonenal (HNE) modifications in RA and associate them with increased levels of autoantibodies in RA patients.

Serum samples from 155 female patients [60 with RA, 35 with osteoarthritis (OA), and 60 healthy controls (HCs)] were obtained. Four novel differential HNE-modified peptide adducts, complement factor H (CFAH)^1211–1230, haptoglobin (HPT)^78–108, immunoglobulin (Ig) kappa chain C region (IGKC)^2–19, and prothrombin (THRB)^328–345, were re-analyzed using tandem mass spectrometric (MS/MS) spectra (ProteomeXchange: PXD004546) from RA patients vs. HCs. Further, we determined serum protein levels of CFAH, HPT, IGKC and THRB, HNE-protein adducts, and autoantibodies against unmodified and HNE-modified peptides. Significant correlations and odds ratios (ORs) were calculated.

Levels of HPT in RA patients were greatly higher than the levels in HCs. Levels of HNE-protein adducts and autoantibodies in RA patients were significantly greater than those of HCs. IgM anti-HPT⁷⁸⁻¹⁰⁸ HNE, IgM anti-IGKC²⁻¹⁹, and IgM anti-IGKC²⁻¹⁹ HNE may be considered as diagnostic biomarkers for RA. Importantly, elevated levels of IgM anti-HPT⁷⁸⁻¹⁰⁸ HNE, IgM anti-IGKC²⁻¹⁹, and IgG anti-THRB^328−345 were positively correlated with the disease activity score in 28 joints for C-reactive protein (DAS28-CRP). Further, the ORs of RA development through IgM anti-HPT⁷⁸⁻¹⁰⁸ HNE (OR 5.235, p < 0.001), IgM anti-IGKC²⁻¹⁹ (OR 12.655, p < 0.001), and IgG anti-THRB^328−345 (OR 5.761, p < 0.001) showed an increased risk. Lastly, we incorporated three machine learning models to differentiate RA from HC and OA, and performed feature selection to determine discriminative features. Experimental results showed that our proposed method achieved an area under the receiver operating characteristic curve of 0.92, which demonstrated that our selected autoantibodies combined with machine learning can efficiently detect RA.

This study discovered that some IgG- and IgM-NAAs and anti-HNE M-NAAs may be correlated with inflammation and disease activity in RA. Moreover, our findings suggested that IgM anti-HPT⁷⁸⁻¹⁰⁸ HNE, IgM anti-IGKC²⁻¹⁹, and IgG anti-THRB^328−345 may play heavy roles in RA development.