Reactivity in ELISA with DNA-loaded nucleosomes in patients with proliferative lupus nephritis

• Both nucleosomes and dsDNA are nephritogenic targets in lupus nephritis. Therefore, we compared in patients with proliferative lupus nephritis the reactivity in ELISA towards dsDNA-loaded nucleosomes with the reactivity towards dsDNA or nucleosomes alone. The sensitivity of the DNA-loaded nucleosome and the nucleosome ELISA was equal (86 and 85%) and higher than the dsDNA ELISA (66%). In patients with absent (dsDNA) or low (nucleosome) reactivity, higher titers were found towards DNA-loaded nucleosomes. Therefore, DNA-loaded nucleosomes seem to provide the best antigenic platform for the evaluation of anti-chromatin antibodies in lupus nephritis.

Autoantibodies against nucleosomes are considered a hallmark of systemic lupus erythematosus (SLE). We compared in patients with proliferative lupus nephritis the diagnostic usefulness of a dsDNA-loaded nucleosome ELISA (anti-dsDNA-NcX) with ELISAs in which dsDNA or nucleosomes alone were coated. First, we analysed whether DNA loading on nucleosomes led to masking of epitopes by using defined monoclonal anti-DNA, anti-histone and nucleosome-specific autoantibodies to evaluate the accessibility of nucleosomal epitopes in the anti-dsDNA-NcX ELISA. Second, autoantibody levels were measured in these 3 ELISAs in 100 patients with proliferative lupus nephritis (LN) before immunosuppressive treatment and in 128 non-SLE disease controls. In patients with LN inter-assay comparisons and associations with clinical and serological parameters were analysed. The panel of monoclonal antibodies revealed that all epitopes were equally accessible in the anti-dsDNA-NcX ELISA as in the two other ELISAs. Patients with proliferative lupus nephritis were positive with dsDNA-loaded nucleosomes in 86%, with DNA in 66% and with nucleosomes in 85%. In the non-lupus disease control group these frequencies were 1.6% (2 out of 128) for both the anti-dsDNA-NcX and the anti-dsDNA ELISA and 0% in the anti-nucleosome ELISA. The levels in the anti-dsDNA-NcX ELISA were high in a group of patients with LN that showed absent reactivity in the anti-DNA or low levels in the anti-nucleosome ELISA. Anti-dsDNA-NcX positivity was associated with higher SLEDAI scores within this group. Within nucleosome-based ELISAs, we propose the anti-dsDNA-NcX ELISA as the preferred test system.