Background/Purpose: Systemic lupus erythematosus (SLE) and lupus nephritis (LN) are autoimmune diseases characterized by circulating antibodies to nuclear self-antigens, including reactivities to double-stranded DNA, RNP and Sm.  Preclinical mouse models exist that mimic aspects of human SLE/LN disease, and are used to study pathogenic mechanisms as well as to test responses to anti-inflammatory treatments.  In clinical samples, autoantibody reactivity to nuclear antigens is heterogeneous, with individual patients exhibiting unique anti-nuclear antibody (ANA) signatures.  We postulated that each distinct mouse model of lupus may also exhibit its own ANA signature, and that by identifying ANA reactivity profiles in preclinical lupus models, robust preclinical biomarker strategies and hypotheses for links with particular aspects of human disease may be devised.  

Methods: A Genalyte Maverick instrument was used to assess IgG reactivity to 13 clinically-relevant nuclear antigens including: SS-A 60, SS-A 52, SS-B, Sm, Sm/RNP, Scl-70, Jo-1, nucleosome, PCNA, Ku, Centromere A & B, Ribosomal P and dsDNA in a simultaneous manner.  Analysis was performed on frozen plasma samples archived from several murine lupus models, including spontaneous NZBW-F1, IFNα-accelerated NZBW-F1, and spontaneous MRL-lpr.  In some samples, both Maverick technology and in-house ELISA assays were used to assess ANA to dsDNA and Sm/RNP for cross-methodology validation.   

Results: Maverick analysis of nuclear antigen reactivities showed that each murine model had a distinct ANA signature.  Spontaneous NZBW-F1 mice developed strong reactivity to dsDNA, with a lesser anti-RNP component.  However, when NZBW-F1 disease was accelerated via injection of a non-replicative IFNα-inducing adenovirus, ANA reactivity was stronger to RNP nuclear antigen, with a lesser anti-dsDNA component.  Both male and female MRL-lpr mice showed strong, age-dependent increases in multiple ANAs including reactivity to dsDNA, RNP, Sm, and nucleosome.  Maverick assessment of ANA reactivities to dsDNA and Sm/RNP significantly correlated to titers generated by in-house ELISAs.  Additional Maverick assessments of ANA from IFNα-accelerated NZBW-F1 mice treated prophylactically with mycophenolate mofetil (Cellcept), showed Cellcept significantly prevented anti-RNP autoantibody production, but only a trend was seen for decreases in anti-dsDNA autoantibody production.  

Conclusion: These results show that each murine lupus model may exhibit its own unique ANA signature, and that Genalyte Maverick technology is a quick and useful methodology for identifying this signature via simultaneous assessment of several ANA from a single plasma sample.  Furthermore, understanding ANA reactivity profiles in each model may help guide better preclinical biomarker design and will have impact on interpreting efficacy of anti-inflammatory treatments.

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ACR Meeting Abstracts - https://acrabstracts.org/abstract/characterization-of-anti-nuclear-antibody-ana-signatures-in-murine-models-of-lupus-using-genalyte-maverick-technology/