Serum IgG2 antibody multi-composition in systemic lupus erythematosus and in lupus nephritis (Part 2): prospective study

Maurizio Bruschi, Gabriella Moroni, Renato Alberto Sinico, Franco Franceschini, Micaela Fredi, Augusto Vaglio, Lorenzo Cavagna, Andrea Petretto, Federico Pratesi, Paola Migliorini, Francesco Locatelli, Giulia Pazzola, Giampaola Pesce, Marcello Bagnasco, Angelo Manfredi, Giuseppe A Ramirez, Pasquale Esposito, Giuseppe Murdaca, Simone Negrini, Leda Cipriani, Barbara Trezzi, Giacomo Emmi, Ilaria Cavazzana, Valentina Binda, Matteo d'Alessandro, Paride Fenaroli, Isabella Pisani, Giacomo Garibotto, Carlomaurizio Montecucco, Domenico Santoro, Francesco Scolari, Stefano Volpi, Marta Mosca, Angela Tincani, Giovanni Candiano, Marco Prunotto, Enrico Verrina, Andrea Angeletti, Angelo Ravelli, Gian Marco Ghiggeri, Maurizio Bruschi, Gabriella Moroni, Renato Alberto Sinico, Franco Franceschini, Micaela Fredi, Augusto Vaglio, Lorenzo Cavagna, Andrea Petretto, Federico Pratesi, Paola Migliorini, Francesco Locatelli, Giulia Pazzola, Giampaola Pesce, Marcello Bagnasco, Angelo Manfredi, Giuseppe A Ramirez, Pasquale Esposito, Giuseppe Murdaca, Simone Negrini, Leda Cipriani, Barbara Trezzi, Giacomo Emmi, Ilaria Cavazzana, Valentina Binda, Matteo d'Alessandro, Paride Fenaroli, Isabella Pisani, Giacomo Garibotto, Carlomaurizio Montecucco, Domenico Santoro, Francesco Scolari, Stefano Volpi, Marta Mosca, Angela Tincani, Giovanni Candiano, Marco Prunotto, Enrico Verrina, Andrea Angeletti, Angelo Ravelli, Gian Marco Ghiggeri

Abstract

Objectives: Circulating anti-ENO1 and anti-H2A IgG2 have been identified as specific signatures of LN in a cross-over approach. We sought to show whether the same antibodies identify selected population of patients with LN with potentially different clinical outcomes.

Methods: Here we report the prospective analysis over 36 months of circulating IgG2 levels in patients with newly diagnosed LN (n=91) and SLE (n=31) and in other patients with SLE recruited within 2 years from diagnosis (n=99). Anti-podocyte (ENO1), anti-nucleosome (DNA, histone 2 A, histone 3) and anti-circulating proteins (C1q, AnnexinA1-ANXA1) IgG2 antibodies were determined by home-made techniques.

Results: LN patients were the main focus of the study. Anti-ENO1, anti-H2A and anti-ANXA1 IgG2 decreased in parallel to proteinuria and normalized within 12 months in the majority of patients while anti-dsDNA IgG2 remained high over the 36 months. Anti-ENO1 and anti-H2A had the highest association with proteinuria (Heat Map) and identified the highest number of patients with high proteinuria (68% and 71% respectively) and/or with reduced estimated glomerula filtration rate (eGFR) (58% for both antibodies) compared with 23% and 17% of anti-dsDNA (agreement analysis). Anti-ENO1 positive LN patients had higher proteinuria than negative patients at T0 and presented the maximal decrement within 12 months.

Conclusions: Anti-ENO1, anti-H2A and anti-ANXA1 antibodies were associated with high proteinuria in LN patients and Anti-ENO1 also presented the maximal reduction within 12 months that paralleled the decrease of proteinuria. Anti-dsDNA were not associated with renal outcome parameters. New IgG2 antibody signatures should be utilized as tracers of personalized therapies in LN.

Trial registration: The Zeus study was registered at https://ichgcp.net/clinical-trials-registry/NCT02403115" title="See in ClinicalTrials.gov">NCT02403115).

Keywords: anti-C1q antibodies; anti-ENO1 antibodies; anti-Histone 2A antibodies; biomarkers; lupus nephritis; systemic lupus erythematosus.

© The Author(s) 2020. Published by Oxford University Press on behalf of the British Society for Rheumatology.

Figures

Fig . 1
Fig. 1
Circulating levels of anti-dsDNA, anti-histone 2 A and anti-histone 3 IgG2 Circulating levels of anti-dsDNA, anti-histone 2A and anti-histone 3 IgG2 were determined in a cohort of 91 LN patients recruited at the time of the diagnosis and were then followed for 36 months. In all cases, antibodies were of the IgG2 isotype and levels were calculated as Relative Intensity value (RU/ml) given the absence of WHO international standards. * indicates a statistical difference with T0. WHO: World Health Organisation.
Fig . 2
Fig. 2
Circulating levels of anti-ENO1, anti-annexin A1 and anti-C1q IgG2 Circulating levels of anti-ENO1, anti-ANXA1 IgG2 and anti-C1q IgG2 were determined in the same group of LN patients of Fig. 1. In all cases, antibodies were of the IgG2 isotype and levels were calculated as Relative Intensity value (RU/ml) given the absence of WHO international standards .* indicates a statistical difference with T0. WHO: World Health Organisation.
Fig . 3
Fig. 3
Variations of circulating antibodies within 12 months of follow-up in LN patients Variations of circulating levels of each antibody from T0 were determined after 12 months of follow-up in LN patients. The decrement of proteinuria is reported for comparison. It is shown that anti-ENO1 and anti-ANXA1 had the maximal decrement and that anti-dsDNA had no variation at all. Results are shown as median and interquartile range. *indicates a statistical difference between anti-dsDNA and anti-ANXA1 with anti-ENO1.
Fig . 4
Fig. 4
Time course of proteinuria and of circulating levels of antibodies in LN patients Circulating levels of each antibody were determined every 12 months for 3 years in LN patients. Results are reported together with proteinuria determined at the same time points. Statistical significance of the decrement in circulating antibodies is reported in Figs 1–3. Statistical significance of proteinuria decrements is reported in Supplementary Fig. S1, available at Rheumatology online.
https://www.ncbi.nlm.nih.gov/pmc/articles/instance/8516512/bin/keaa793f5.jpg
Fig. 5 Proteinuria at T0 and modifications during the follow-up in anti-ENO1 positive and negative LN patients Proteinuria at T0 and % changes during the first 12 months of follow-up are reported for patients positive and negative for anti-ENO1. In the former case (proteinuria levels) a statistical difference was observed between the two groups. Also, the decrement of proteinuria was more evident in anti-ENO1 positive patients. The same parameters are reported sub-dividing the whole LN group in respect to positivity for anti-dsDNA IgG2.

References

    1. Rahman A, Isenberg DA.. Systemic lupus erythematosus. N Engl J Med 2008;358:929–39.
    1. Bagavant H, Fu SM.. Pathogenesis of kidney disease in systemic lupus erythematosus. Curr Opin Rheumatol 2009;21:489–94.
    1. Hanly JG, O’Keeffe AG, Su L. et al. The frequency and outcome of lupus nephritis: results from an international inception cohort study. Rheumatology 2016;55:252–62.
    1. Mjelle JE, Mjelle JE, Kalaaji M. et al. Exposure of chromatin and not high affinity for dsDNA determines the nephritogenic impact of anti-dsDNA antibodies in (NZBxNZW)F1 mice. Autoimmunity 2009;42:104–11.
    1. Xie C, Liang Z, Chang S, Mohan C.. Use of a novel elution regimen reveals the dominance of polyreactive antinuclear autoantibodies in lupus kidneys. Arthritis Rheum 2003;48:2343–52.
    1. Kalaaji M, Sturfelt G, Mjelle JE, Nossent H, Rekvig OP.. Critical comparative analyses of anti-alpha-actinin and glomerulus-bound antibodies in human and murine lupus nephritis. Arthritis Rheum 2006;54:914–26.
    1. Kalaaji M, Mortensen E, Jorgensen L, Olsen R, Rekvig OP.. Nephritogenic lupus antibodies recognize glomerular basement membrane-associated chromatin fragments released from apoptotic intraglomerular cells. Am J Pathol 2006;168:1779–92.
    1. Yung S, Cheung KF, Zhang Q, Chan TM.. Anti-dsDNA antibodies bind to mesangial annexin II in lupus nephritis. J Am Soc Nephrol 2010;21:1912–27.
    1. Romay-Penabad Z, Montiel-Manzano MG, Shilagard T. et al. Annexin A2 is involved in antiphospholipid antibody-mediated pathogenic effects in vitro and in vivo. Blood 2009;114:3074–83.
    1. Krishnan MR, Wang C, Marion TN.. Anti-DNA autoantibodies initiate experimental lupus nephritis by binding directly to the glomerular basement membrane in mice. Kidney Int 2012;82:184–92.
    1. Rekvig OP. The anti-DNA antibody: origin and impact, dogmas and controversies. Nat Rev Rheumatol 2015;11:530–40.
    1. Font J, Cervera R, Ramos-Casals M. et al. Clusters of clinical and immunologic features in systemic lupus erythematosus: analysis of 600 patients from a single center. Semin Arthritis Rheum 2004;33:217–30.
    1. Gensous N, Marti A, Barnetche T, on behalf of the FHU ACRONIM et al. Predictive biological markers of systemic lupus erythematosus flares: a systematic literature review. Arthritis Res Ther 2017;19:238.
    1. Yang J, Xu Z, Sui M. et al. Co-positivity for anti-dsDNA, -nucleosome and -histone antibodies in lupus nephritis is indicative of high serum levels and severe nephropathy. PLoS One 2015;10:e0140441.
    1. Kavanaugh AF, Solomon DH, The American College of Rheumatology Ad Hoc Committee on Immunologic Testing Guidelines. Guidelines for immunologic laboratory testing in the rheumatic diseases: anti-DNA antibody tests. Arthritis Rheum 2002;47:546–55.
    1. Moroni G, Radice A, Giammarresi G. et al. Are laboratory tests useful for monitoring the activity of lupus nephritis? A 6-year prospective study in a cohort of 228 patients with lupus nephritis. Ann Rheum Dis 2009;68:234–7.
    1. Sinico RA, Rimoldi L, Radice A. et al. Anti-C1q autoantibodies in lupus nephritis. Ann N Y Acad Sci 2009;1173:47–51.
    1. Mannik M, Merrill CE, Stamps LD, Wener MH.. Multiple autoantibodies form the glomerular immune deposits in patients with systemic lupus erythematosus. J Rheumatol 2003;30:1495–504.
    1. Bruschi M, Galetti M, Sinico RA. et al. Glomerular autoimmune multicomponents of human lupus nephritis in vivo (2): planted antigens. J Am Soc Nephrol 2015;26:1905–24.
    1. Bonanni A, Vaglio A, Bruschi M. et al. Multi-antibody composition in lupus nephritis: isotype and antigen specificity make the difference. Autoimmun Rev 2015;14:692–702.
    1. Petri M, Orbai AM, Alarcon GS. et al. Derivation and validation of the Systemic Lupus International Collaborating Clinics classification criteria for systemic lupus erythematosus. Arthritis Rheum 2012;64:2677–86.
    1. Moroni G, Vercelloni PG, Quaglini S. et al. Changing patterns in clinical-histological presentation and renal outcome over the last five decades in a cohort of 499 patients with lupus nephritis. Ann Rheum Dis 2018;77:1318–25.
    1. Zweig MH, Campbell G.. Receiver-operating characteristic (ROC) plots: a fundamental evaluation tool in clinical medicine. Clin Chem 1993;39:561–77.
    1. Bruschi M, Petretto A, Vaglio A. et al. Annexin A1 and autoimmunity: from basic science to clinical applications. Int J Mol Sci 2018;19.
    1. Bruschi M, Petretto A, Santucci L. et al. Neutrophil Extracellular Traps protein composition is specific for patients with Lupus nephritis and includes methyl-oxidized alphaenolase (methionine sulfoxide 93). Sci Rep 2019;9:7934.
    1. Dieker J, Tel J, Pieterse E. et al. Circulating apoptotic microparticles in systemic lupus erythematosus patients drive the activation of dendritic cell subsets and prime neutrophils for NETosis. Arthritis Rheumatol 2016;68:462–72.
    1. Ullal AJ, Reich CF 3rd, Clowse M. et al. Microparticles as antigenic targets of antibodies to DNA and nucleosomes in systemic lupus erythematosus. J Autoimmun 2011;36:173–80.
    1. Napirei M, Ludwig S, Mezrhab J, Klöckl T, Mannherz HG.. Murine serum nucleases–contrasting effects of plasmin and heparin on the activities of DNase1 and DNase1-like 3 (DNase1l3). FEBS J 2009;276:1059–73.
    1. Sisirak V, Sally B, D’Agati V. et al. Digestion of chromatin in apoptotic cell microparticles prevents autoimmunity. Cell 2016;166:88–101.
    1. Bruschi M, Petretto A, Bertelli R. et al. Post-translational modified proteins are biomarkers of autoimmune-processes: NETosis and the inflammatory-autoimmunity connection. Clin Chim Acta 2017;464:12–6.
    1. Bruschi M, Bonanni A, Petretto A. et al. Neutrophil Extracellular Traps (NETs) profiles in patients with incident SLE and lupus nephritis. J Rheumatol 2020;47:377–86.
    1. Lood C, Blanco LP, Purmalek MM. et al. Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease. Nat Med 2016;22:146–53.
    1. Muller S, Radic M.. Oxidation and mitochondrial origin of NET DNA in the pathogenesis of lupus. Nat Med 2016;22:126–7.

Source: PubMed

Szponzorok és közreműködők

Egészségi állapot

Kábítószer-beavatkozások

3
Iratkozz fel