Neutrophil Extracellular Traps in the Autoimmunity Context

Maurizio Bruschi, Gabriella Moroni, Renato Alberto Sinico, Franco Franceschini, Micaela Fredi, Augusto Vaglio, Lorenzo Cavagna, Andrea Petretto, Federico Pratesi, Paola Migliorini, Angelo Manfredi, Giuseppe A Ramirez, Pasquale Esposito, Simone Negrini, Barbara Trezzi, Giacomo Emmi, 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, Angelo Manfredi, Giuseppe A Ramirez, Pasquale Esposito, Simone Negrini, Barbara Trezzi, Giacomo Emmi, Domenico Santoro, Francesco Scolari, Stefano Volpi, Marta Mosca, Angela Tincani, Giovanni Candiano, Marco Prunotto, Enrico Verrina, Andrea Angeletti, Angelo Ravelli, Gian Marco Ghiggeri

Abstract

The formation of neutrophil extracellular traps (NETs) is a strategy utilized by neutrophils for capturing infective agents. Extracellular traps consist in a physical net made of DNA and intracellular proteins externalized from neutrophils, where bacteria and viruses are entrapped and killed by proteolysis. A complex series of events contributes to achieving NET formation: signaling from infectious triggers comes first, followed by decondensation of chromatin and extrusion of the nucleosome components (DNA, histones) from the nucleus and, after cell membrane breakdown, outside the cell. NETs are composed of either DNA or nucleosome proteins and hundreds of cytoplasm proteins, a part of which undergo post-translational modification during the steps leading to NETs. There is a thin balance between the production and the removal of circulating NETs from blood where digestion of DNA by circulating DNases 1 and IL3 has a critical role. A delay in NET removal may have consequences for autoimmunity. Recent studies have shown that circulating NET levels are increased in systemic lupus erythematosus (SLE) for a functional block of NET removal mediated by anti-DNase antibodies or, in rare cases, by DNase IL3 mutations. In SLE, the persistence in circulation of NETs signifies elevated concentrations of either free DNA/nucleosome components and oxidized proteins that, in some cases, are recognized as non-self and presented to B-cells by Toll-like receptor 9 (TLR9). In this way, it is activated as an immunologic response, leading to the formation of IgG2 auto-antibody. Monitoring serum NET levels represents a potential new way to herald the development of renal lesions and has clinical implications. Modulating the balance between NET formation and removal is one of the objectives of basic research that are aimed to design new drugs for SLE. Clinical Trial Registration Number: The Zeus study was registered at https://ichgcp.net/clinical-trials-registry/NCT02403115" title="See in ClinicalTrials.gov">NCT02403115).

Keywords: Lupus nephritis; anti-C1q antibodies; anti-alpha enolase; anti-histone; biomarker; systemic lupu erythematosus.

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2021 Bruschi, Moroni, Sinico, Franceschini, Fredi, Vaglio, Cavagna, Petretto, Pratesi, Migliorini, Manfredi, Ramirez, Esposito, Negrini, Trezzi, Emmi, Santoro, Scolari, Volpi, Mosca, Tincani, Candiano, Prunotto, Verrina, Angeletti, Ravelli and Ghiggeri.

Figures

Figure 1
Figure 1
Traditional immunofluorescence microscopy analysis of neutrophil extracellular trap (NET) filaments showing that αenolase and DNA are intensely present in NET filaments and co-localized in large segments. The images were acquired using LSM 510 Meta confocal system scan.
Figure 2
Figure 2
Schematic representation of the pathway that potentially connects neutrophil extracellular traps (NETs) with the formation of antibodies of IgG2 isotype in systemic lupus erythematosus. The various steps are detailed in the text. As for the last step that involves TLR9, the data supporting a direct link between NETs and IgG2 via TLR9 are presented in this special issue of Frontiers (The kidney in inflammatory and immune-mediated diseases) (see Bertelli et al.).

References

    1. Ghiggeri GM, D'Alessandro M, Bartolomeo D, Degl'Innocenti ML, Magnasco A, Lugani F, et al. . An update on antibodies to necleosome components as biomarkers of sistemic Lupus erythematosus and of Lupus flares. Int J Mol Sci. (2019) 20:22. 10.3390/ijms20225799
    1. Bruschi M, Petretto A, Santucci L, Vaglio A, Pratesi F, Migliorini P, 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. 10.1038/s41598-019-44379-w
    1. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, et al. . Neutrophil extracellular traps kill bacteria. Science. (2004) 303:1532–5. 10.1126/science.1092385
    1. Nathan C. Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol. (2006) 6:173–82. 10.1038/nri1785
    1. Fuchs TA, Abed U, Goosmann C, Hurwitz R, Schulze I, Wahn V, et al. . Novel cell death program leads to neutrophil extracellular traps. J Cell Biol. (2007) 176:231–41. 10.1083/jcb.200606027
    1. Urban C, Zychlinsky A. Netting bacteria in sepsis. Nat Med. (2007) 13:403–4. 10.1038/nm0407-403
    1. Bianchi M, Hakkim A, Brinkmann V, Siler U, Seger RA, Zychlinsky A, et al. . Restoration of NET formation by gene therapy in CGD controls aspergillosis. Blood. (2009) 114:2619–22. 10.1182/blood-2009-05-221606
    1. Steinberg BE, Grinstein S. Unconventional roles of the NADPH oxidase: signaling, ion homeostasis, and cell death. Sci STKE. (2007) 2007:pe11. 10.1126/stke.3792007pe11
    1. Maitra U, Singh N, Gan L, Ringwood L, Li L. IRAK-1 contributes to lipopolysaccharide-induced reactive oxygen species generation in macrophages by inducing NOX-1 transcription and Rac1 activation and suppressing the expression of antioxidative enzymes. J Biol Chem. (2009) 284:35403–11. 10.1074/jbc.M109.059501
    1. Kim JS, Yeo S, Shin DG, Bae YS, Lee JJ, Chin BR, et al. . Glycogen synthase kinase 3beta and beta- catenin pathway is involved in toll-like receptor 4-mediated NADPH oxidase 1 expression in macrophages. FEBS J. (2010) 277:2830–7. 10.1111/j.1742-4658.2010.07700.x
    1. Hakkim A, Fuchs TA, Martinez NE, Hess S, Prinz H, Zychlinsky A, et al. . Activation of the Raf-MEK-ERK pathway is required for neutrophil extracellular trap formation. Nat Chem Biol. (2011) 7:75–7. 10.1038/nchembio.496
    1. Douda DN, Khan MA, Grasemann H, Palaniyar N. SK3 channel and mitochondrial ROS mediate NADPH oxidase-independent NETosis induced by calcium influx. Proc Natl Acad Sci USA. (2015) 112:2817–22. 10.1073/pnas.1414055112
    1. Keshari RS, Verma A, Barthwal MK, Dikshit M. Reactive oxygen species-induced activation of ERK and p38 MAPK mediates PMA-induced NETs release from human neutrophils. J Cell Biochem. (2013) 114:532–40. 10.1002/jcb.24391
    1. Garcia-Romo GS, Caielli S, Vega B, Connolly J, Allantaz F, Xu Z, et al. . Netting neutrophils are major inducers of type I IFN production in pediatric systemic lupus erythematosus. Sci Transl Med. (2011) 3:73ra20. 10.1126/scitranslmed.3001201
    1. Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa U, Nishizuka Y. Direct activation of calcium- activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. (1982) 257:7847–51. 10.1016/S0021-9258(18)34459-4
    1. Bouin AP, Grandvaux N, Vignais PV, Fuchs A. p40(phox) is phosphorylated on threonine 154 and serine 315 during activation of the phagocyte NADPH oxidase. Implication of a protein kinase c-type kinase in the phosphorylation process. J Biol Chem. (1998) 273:30097–103. 10.1074/jbc.273.46.30097
    1. Metzler KD, Fuchs TA, Nauseef WM, Reumaux D, Roesler J, Schulze I, et al. . Myeloperoxidase is required for neutrophil extracellular trap formation: implications for innate immunity. Blood. (2011) 117:953–9. 10.1182/blood-2010-06-290171
    1. Metzler KD, Goosmann C, Lubojemska A, Zychlinsky A, Papayannopoulos V. A myeloperoxidase- containing complex regulates neutrophil elastase release and actin dynamics during NETosis. Cell Rep. (2014) 8:883–96. 10.1016/j.celrep.2014.06.044
    1. Wang Y, Li M, Stadler S, Correll S, Li P, Wang D, et al. . Histone hypercitrullination mediates chromatin decondensation and neutrophil extracellular trap formation. J Cell Biol. (2009) 184:205–13. 10.1083/jcb.200806072
    1. Li P, Li M, Lindberg MR, Kennett MJ, Xiong N, Wang Y. PAD4 is essential for antibacterial innate immunity mediated by neutrophil extracellular traps. J Exp Med. (2010) 207:1853–62. 10.1084/jem.20100239
    1. Ullal AJ, Reich CF, III, Clowse M, Criscione-Schreiber LG, Tochacek M, Monestier M, et al. . Microparticles as antigenic targets of antibodies to DNA and nucleosomes in systemic lupus erythematosus. J Autoimmun. (2011) 36:173–80. 10.1016/j.jaut.2011.02.001
    1. Bruschi M, Galetti M, Sinico RA, Moroni G, Bonanni A, Radice A, et al. . Glomerular autoimmune multicomponents of human lupus nephritis in vivo (2): planted antigens. J Am Soc Nephrol. (2015) 26:1905–24. 10.1681/ASN.2014050493
    1. Bruschi M, Sinico RA, Moroni G, Pratesi F, Migliorini P, Galetti M, et al. . Glomerular autoimmune multicomponents of human lupus nephritis in vivo: alpha-enolase and annexin AI. J Am Soc Nephrol. (2014) 25:2483–98. 10.1681/ASN.2013090987
    1. Napirei M, Ludwig S, Mezrhab J, Klockl 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. 10.1111/j.1742-4658.2008.06849.x
    1. Napirei M, Karsunky H, Zevnik B, Stephan H, Mannherz HG, Moroy T. Features of systemic lupus erythematosus in Dnase1-deficient mice. Nat Genet. (2000) 25:177–81. 10.1038/76032
    1. Sisirak V, Sally B, D'Agati V, Martinez-Ortiz W, Ozcakar ZB, David J, et al. . Digestion of chromatin in apoptotic cell microparticles prevents autoimmunity. Cell. (2016) 166:88–101. 10.1016/j.cell.2016.05.034
    1. Martin M, Leffler J, Blom AM. Annexin A2 and A5 serve as new ligands for C1q on apoptotic cells. J Biol Chem. (2012) 287:33733–44. 10.1074/jbc.M112.341339
    1. Urban CF, Ermert D, Schmid M, Abu-Abed U, Goosmann C, Nacken W, et al. . Neutrophil extracellular traps contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans. PLoS Pathog. (2009) 5:e1000639. 10.1371/journal.ppat.1000639
    1. Petretto A, Bruschi M, Pratesi F, Croia C, Candiano G, Ghiggeri G, et al. . Neutrophil extracellular traps (NET) induced by different stimuli: a comparative proteomic analysis. PLoS ONE. (2019) 14:e0218946. 10.1371/journal.pone.0218946
    1. Pilsczek FH, Salina D, Poon KK, Fahey C, Yipp BG, Sibley CD, et al. . A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus. J Immunol. (2010) 185:7413–25. 10.4049/jimmunol.1000675
    1. Yipp BG, Kubes P. NETosis: how vital is it? Blood. (2013) 122:2784–94. 10.1182/blood-2013-04-457671
    1. Romero V, Fert-Bober J, Nigrovic PA, Darrah E, Haque UJ, Lee DM, et al. . Immune-mediated pore- forming pathways induce cellular hypercitrullination and generate citrullinated autoantigens in rheumatoid arthritis. Sci Transl Med. (2013) 5:209ra150. 10.1126/scitranslmed.3006869
    1. Gupta S, Kaplan MJ. The role of neutrophils and NETosis in autoimmune and renal diseases. Nat Rev Nephrol. (2016) 12:402–13. 10.1038/nrneph.2016.71
    1. Bonanni A, Vaglio A, Bruschi M, Sinico RA, Cavagna L, Moroni G, et al. . Multi-antibody composition in lupus nephritis: isotype and antigen specificity make the difference. Autoimmun Rev. (2015) 14:692–702. 10.1016/j.autrev.2015.04.004
    1. Bruschi M, Moroni G, Sinico RA, Franceschini F, Fredi M, Vaglio A, et al. . Serum IgG2 antibody multicomposition in systemic lupus erythematosus and lupus nephritis (Part 1): cross-sectional analysis. Rheumatology. (2020). 10.1093/rheumatology/keaa767. [Epub ahead of print].
    1. Bruschi M, Moroni G, Sinico RA, Franceschini F, Fredi M, Vaglio A, et al. . Serum IgG2 antibody multi- composition in systemic lupus erythematosus and in lupus nephritis (Part 2): prospective study. Rheumatology. (2020). 10.1093/rheumatology/keaa793. [Epub ahead of print].
    1. Bruschi M, Petretto A, Bertelli R, Galetti M, Bonanni A, Pratesi F, et al. . Post-translational modified proteins are biomarkers of autoimmune-processes: NETosis and the inflammatory-autoimmunity connection. Clin Chim Acta. (2017) 464:12–6. 10.1016/j.cca.2016.11.006
    1. Denny MF, Yalavarthi S, Zhao W, Thacker SG, Anderson M, Sandy AR, et al. . A distinct subset of proinflammatory neutrophils isolated from patients with systemic lupus erythematosus induces vascular damage and synthesizes type I IFNs. J Immunol. (2010) 184:3284–97. 10.4049/jimmunol.0902199
    1. Villanueva E, Yalavarthi S, Berthier CC, Hodgin JB, Khandpur R, Lin AM, et al. . Netting neutrophils induce endothelial damage, infiltrate tissues, and expose immunostimulatory molecules in systemic lupus erythematosus. J Immunol. (2011) 187:538–52. 10.4049/jimmunol.1100450
    1. Hakkim A, Furnrohr BG, Amann K, Laube B, Abed UA, Brinkmann V, et al. . Impairment of neutrophil extracellular trap degradation is associated with lupus nephritis. Proc Natl Acad Sci USA. (2010) 107:9813–8. 10.1073/pnas.0909927107
    1. Kessenbrock K, Krumbholz M, Schonermarck U, Back W, Gross WL, Werb Z, et al. . Netting neutrophils in autoimmune small-vessel vasculitis. Nat Med. (2009) 15:623–5. 10.1038/nm.1959
    1. Soderberg D, Kurz T, Motamedi A, Hellmark T, Eriksson P, Segelmark M. Increased levels of neutrophil extracellular trap remnants in the circulation of patients with small vessel vasculitis, but an inverse correlation to anti-neutrophil cytoplasmic antibodies during remission. Rheumatology. (2015) 54:2085–94. 10.1093/rheumatology/kev217
    1. Knight JS, Zhao W, Luo W, Subramanian V, O'Dell AA, Yalavarthi S, et al. . Peptidylarginine deiminase inhibition is immunomodulatory and vasculoprotective in murine lupus. J Clin Invest. (2013) 123:2981–93. 10.1172/JCI67390
    1. Zhang S, Lu X, Shu X, Tian X, Yang H, Yang W, et al. . Elevated plasma cfDNA may be associated with active lupus nephritis and partially attributed to abnormal regulation of neutrophil extracellular traps (NETs) in patients with systemic lupus erythematosus. Intern Med. (2014) 53:2763–71. 10.2169/internalmedicine.53.2570
    1. Lood C, Blanco LP, Purmalek MM, Carmona-Rivera C, De Ravin SS, Smith CK, et al. . Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease. Nat Med. (2016) 22:146–53. 10.1038/nm.4027
    1. Bruschi M, Bonanni A, Petretto A, Vaglio A, Pratesi F, Santucci L, et al. . Neutrophil Extracellular Traps (NETs) profiles in patients with incident SLE and lupus nephritis. J Rheumatol. (2019) 47:377–386. 10.3899/jrheum.181232
    1. Zykova SN, Tveita AA, Rekvig OP. Renal Dnase1 enzyme activity and protein expression is selectively shut down in murine and human membranoproliferative lupus nephritis. PLoS ONE. (2010) 5:e0012096. 10.1371/journal.pone.0012096
    1. Ozcakar ZB, Foster J, 2nd, Diaz-Horta O, Kasapcopur O, Fan YS, Yalcinkaya F, et al. . DNASE1L3 mutations in hypocomplementemic urticarial vasculitis syndrome. Arthritis Rheum. (2013) 65:2183–9. 10.1002/art.38010
    1. Rice GI, Melki I, Fremond ML, Briggs TA, Rodero MP, Kitabayashi N, et al. . Assessment of type I interferon signaling in pediatric inflammatory disease. J Clin Immunol. (2017) 37:123–32. 10.1007/s10875-016-0359-1
    1. Shin HD, Park BL, Kim LH, Lee HS, Kim TY, Bae SC. Common DNase I polymorphism associated with autoantibody production among systemic lupus erythematosus patients. Hum Mol Genet. (2004) 13:2343–50. 10.1093/hmg/ddh275
    1. Yasutomo K, Horiuchi T, Kagami S, Tsukamoto H, Hashimura C, Urushihara M, et al. . Mutation of DNASE1 in people with systemic lupus erythematosus. Nat Genet. (2001) 28:313–4. 10.1038/91070
    1. Fenton K, Fismen S, Hedberg A, Seredkina N, Fenton C, Mortensen ES, et al. . Anti-dsDNA antibodies promote initiation, and acquired loss of renal Dnase1 promotes progression of lupus nephritis in autoimmune (NZBxNZW)F1 mice. PLoS ONE. (2009) 4:e8474. 10.1371/journal.pone.0008474
    1. Seredkina N, Zykova SN, Rekvig OP. Progression of murine lupus nephritis is linked to acquired renal Dnase1 deficiency and not to up-regulated apoptosis. Am J Pathol. (2009) 175:97–106. 10.2353/ajpath.2009.080943
    1. Nakazawa D, Shida H, Kusunoki Y, Miyoshi A, Nishio S, Tomaru U, et al. . The responses of macrophages in interaction with neutrophils that undergo NETosis. J Autoimmun. (2016) 67:19–28. 10.1016/j.jaut.2015.08.018
    1. Garcia GE, Truong LD, Li P, Zhang P, Johnson RJ, Wilson CB, et al. . Inhibition of CXCL16 attenuates inflammatory and progressive phases of anti-glomerular basement membrane antibody-associated glomerulonephritis. Am J Pathol. (2007) 170:1485–96. 10.2353/ajpath.2007.060065
    1. Biron BM, Chung CS, Chen Y, Wilson Z, Fallon EA, Reichner JS, et al. . PAD4 deficiency leads to decreased organ dysfunction and improved survival in a dual insult model of hemorrhagic shock and sepsis. J Immunol. (2018) 200:1817–28. 10.4049/jimmunol.1700639
    1. Sondo E, Bertelli R, Pesce E, Ghiggeri GM, Pedemonte N. High-Content screening identifies vanilloids as a novel class of inhibitors of NET formation. Front Immunol. (2019) 10:963. 10.3389/fimmu.2019.00963
    1. Neeli I, Radic M. Opposition between PKC isoforms regulates histone deimination and neutrophil extracellular chromatin release. Front Immunol. (2013) 4:38. 10.3389/fimmu.2013.00038
    1. Khandpur R, Carmona-Rivera C, Vivekanandan-Giri A, Gizinski A, Yalavarthi S, Knight JS, et al. . NETs are a source of citrullinated autoantigens and stimulate inflammatory responses in rheumatoid arthritis. Sci Transl Med. (2013) 5:178ra40. 10.1126/scitranslmed.3005580
    1. Yang C, Chilvers M, Montgomery M, Nolan SJ. Dornase alfa for cystic fibrosis. Cochrane Database Syst Rev. (2016) 4:CD001127. 10.1002/14651858.CD001127.pub3
    1. Cantin AM. DNase I acutely increases cystic fibrosis sputum elastase activity and its potential to induce lung hemorrhage in mice. Am J Respir Crit Care Med. (1998) 157:464–9. 10.1164/ajrccm.157.2.9608033
    1. Tetz V, Tetz G. Effect of deoxyribonuclease I treatment for dementia in end-stage Alzheimer's disease: a case report. J Med Case Rep. (2016) 10:131. 10.1186/s13256-016-0931-6
    1. Macanovic M, Sinicropi D, Shak S, Baughman S, Thiru S, Lachmann PJ. The treatment of systemic lupus erythematosus (SLE) in NZB/W F1 hybrid mice; studies with recombinant murine DNase and with dexamethasone. Clin Exp Immunol. (1996) 106:243–52. 10.1046/j.1365-2249.1996.d01-839.x
    1. Verthelyi D, Dybdal N, Elias KA, Klinman DM. DNAse treatment does not improve the survival of lupus prone (NZB x NZW)F1 mice. Lupus. (1998) 7:223–30. 10.1191/096120398678920037
    1. Davis JC, Jr, Manzi S, Yarboro C, Rairie J, McInnes I, et al. . Recombinant human Dnase I (rhDNase) in patients with lupus nephritis. Lupus. (1999) 8:68–76. 10.1191/096120399678847380

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