Maurizio Bruschi1,2, Alice Bonanni1,2, Andrea Petretto1,2, Augusto Vaglio1,2, Federico Pratesi1,2, Laura Santucci1,2, Paola Migliorini1,2, Roberta Bertelli1,2, Maricla Galetti1,2, Silvana Belletti1,2, Lorenzo Cavagna1,2, Gabriella Moroni1,2, Franco Franceschini1,2, Micaela Fredi1,2, Giulia Pazzola1,2, Landino Allegri1,2, Renato Alberto Sinico1,2, Giampaola Pesce1,2, Marcello Bagnasco1,2, Angelo Manfredi1,2, Giuseppe A Ramirez1,2, Paola Ramoino1,2, Paolo Bianchini1,2, Francesco Puppo1,2, Francesca Pupo1,2, Simone Negrini1,2, Federico Mattana1,2, Giacomo Emmi1,2, Giacomo Garibotto1,2, Domenico Santoro1,2, Francesco Scolari1,2, Angelo Ravelli1,2, Angela Tincani1,2, Paolo Cravedi1,2, Stefano Volpi1,2, Giovanni Candiano1,2, Gian Marco Ghiggeri3,4. 1. From the Laboratory of Molecular Nephrology, Core Facilities-Proteomics Laboratory, Division of Paediatric Rheumatology and Division of Nephrology, Dialysis, and Transplantation, Scientific Institute for Research and Health Care (IRCCS) Istituto Giannina Gaslini, Genoa; Nephrology Unit, University Hospital, University of Parma; Clinical Immunology Unit, Department of Internal Medicine, University of Pisa; Italian Workers' Compensation Authority (INAIL) Research Center at the University of Parma, Italy; S.Bi.Bi.T Department of the University of Parma, Parma; Division of Rheumatology, University and IRCCS Policlinico S. Matteo, Pavia; Division of Nephrology and Dialysis, IRCCS, Regina Elena, Milan; Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia, Brescia; Nephrology and Dialysis, Arciospedale Santa Maria Nuova, Reggio Emilia; Department of Medicine and Surgery, University of Milan, Bicocca; Medical and Radiometabolic Therapy Unit, Department of Internal Medicine, University of Genoa; Unit of Internal Medicine and Immunology, IRCCS Ospedale San Raffaele, Milan; Department of Earth, Environment and Life Sciences (DISTAV), University of Genoa; Department of Nanophysics, Italian Institute of Technology (IIT); Department of Internal Medicine, University of Genoa; Division of Nephrology, University of Genoa and Policlinico San Martino, Genoa; Lupus Clinic Department of Biomedicine, University of Florence, University Hospital Careggi, Florence; Nephrology and Dialysis Unit, University of Messina and G. Martino Hospital, Messina; Division of Nephrology and Dialysis, University of Brescia and Ospedale di Montichiari, Brescia, Italy; Nephrology Division, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA. 2. M. Bruschi, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; A. Bonanni, MD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; A. Petretto, PhD, Core Facilities-Proteomics Laboratory, IRCCS Istituto Giannina Gaslini; A. Vaglio, MD, Nephrology Unit, University Hospital, University of Parma; F. Pratesi, MD, Clinical Immunology Unit, Department of Internal Medicine, University of Pisa; L. Santucci, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; P. Migliorini, MD, Clinical Immunology Unit, Department of Internal Medicine, University of Pisa; R. Bertelli, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; M. Galetti, PhD, INAIL Research Center at the University of Parma; S. Belletti, MD, S.Bi.Bi.T Department of the University of Parma; L. Cavagna, MD, Division of Rheumatology, University and IRCCS Policlinico S. Matteo; G. Moroni, MD, Division of Nephrology and Dialysis, Fondazione IRCCS Ca' Granda Ospedale Maggiore Milano; F. Franceschini, MD, Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia; M. Fredi, MD, Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia; G. Pazzola, MD, Nephrology and Dialysis, Arciospedale Santa Maria Nuova; L. Allegri, MD, Nephrology Unit, University Hospital, University of Parma; R.A. Sinico, MD, Department of Medicine and Surgery, University of Milan; G. Pesce, PhD, Medical and Radiometabolic Therapy Unit, Department of Internal Medicine, University of Genoa; M. Bagnasco, MD, Medical and Radiometabolic Therapy Unit, Department of Internal Medicine, University of Genoa; A. Manfredi, MD, Unit of Internal Medicine and Immunology, IRCCS Ospedale San Raffaele; G.A. Ramirez, MD, Unit of Internal Medicine and Immunology, IRCCS Ospedale San Raffaele; P. Ramoino, MD, DISTAV, University of Genoa; P. Bianchini, MD, Department of Nanophysics, IIT; F. Puppo, MD, Department of Internal Medicine, University of Genoa; F. Pupo, MD, Department of Internal Medicine, University of Genoa; S. Negrini, MD, Department of Internal Medicine, University of Genoa; F. Mattana, MD, Division of Nephrology, University of Genoa and Policlinico San Martino; G. Emmi, MD, Lupus Clinic Department of Biomedicine, University of Florence, University Hospital Careggi; G. Garibotto, MD, Division of Nephrology, University of Genoa and Policlinico San Martino; D. Santoro, MD, Nephrology and Dialysis Unit, University of Messina and G. Martino Hospital; F. Scolari, MD, Division of Nephrology and Dialysis, University of Brescia and Ospedale di Montichiari; A. Ravelli, MD, Division of Paediatric Rheumatology, IRCCS Istituto Giannina Gaslini; A. Tincani, MD, Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia; P. Cravedi, MD, Nephrology Division, Department of Medicine, Icahn School of Medicine at Mount Sinai; S. Volpi, MD, Division of Paediatric Rheumatology, IRCCS Istituto Giannina Gaslini; G. Candiano, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; G.M. Ghiggeri, MD, PhD, Laboratory of Molecular Nephrology, and Division of Nephrology, Dialysis, and Transplantation, IRCCS Istituto Giannina Gaslini. 3. From the Laboratory of Molecular Nephrology, Core Facilities-Proteomics Laboratory, Division of Paediatric Rheumatology and Division of Nephrology, Dialysis, and Transplantation, Scientific Institute for Research and Health Care (IRCCS) Istituto Giannina Gaslini, Genoa; Nephrology Unit, University Hospital, University of Parma; Clinical Immunology Unit, Department of Internal Medicine, University of Pisa; Italian Workers' Compensation Authority (INAIL) Research Center at the University of Parma, Italy; S.Bi.Bi.T Department of the University of Parma, Parma; Division of Rheumatology, University and IRCCS Policlinico S. Matteo, Pavia; Division of Nephrology and Dialysis, IRCCS, Regina Elena, Milan; Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia, Brescia; Nephrology and Dialysis, Arciospedale Santa Maria Nuova, Reggio Emilia; Department of Medicine and Surgery, University of Milan, Bicocca; Medical and Radiometabolic Therapy Unit, Department of Internal Medicine, University of Genoa; Unit of Internal Medicine and Immunology, IRCCS Ospedale San Raffaele, Milan; Department of Earth, Environment and Life Sciences (DISTAV), University of Genoa; Department of Nanophysics, Italian Institute of Technology (IIT); Department of Internal Medicine, University of Genoa; Division of Nephrology, University of Genoa and Policlinico San Martino, Genoa; Lupus Clinic Department of Biomedicine, University of Florence, University Hospital Careggi, Florence; Nephrology and Dialysis Unit, University of Messina and G. Martino Hospital, Messina; Division of Nephrology and Dialysis, University of Brescia and Ospedale di Montichiari, Brescia, Italy; Nephrology Division, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA. GMarcoGhiggeri@ospedale-gaslini.ge.it. 4. M. Bruschi, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; A. Bonanni, MD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; A. Petretto, PhD, Core Facilities-Proteomics Laboratory, IRCCS Istituto Giannina Gaslini; A. Vaglio, MD, Nephrology Unit, University Hospital, University of Parma; F. Pratesi, MD, Clinical Immunology Unit, Department of Internal Medicine, University of Pisa; L. Santucci, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; P. Migliorini, MD, Clinical Immunology Unit, Department of Internal Medicine, University of Pisa; R. Bertelli, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; M. Galetti, PhD, INAIL Research Center at the University of Parma; S. Belletti, MD, S.Bi.Bi.T Department of the University of Parma; L. Cavagna, MD, Division of Rheumatology, University and IRCCS Policlinico S. Matteo; G. Moroni, MD, Division of Nephrology and Dialysis, Fondazione IRCCS Ca' Granda Ospedale Maggiore Milano; F. Franceschini, MD, Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia; M. Fredi, MD, Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia; G. Pazzola, MD, Nephrology and Dialysis, Arciospedale Santa Maria Nuova; L. Allegri, MD, Nephrology Unit, University Hospital, University of Parma; R.A. Sinico, MD, Department of Medicine and Surgery, University of Milan; G. Pesce, PhD, Medical and Radiometabolic Therapy Unit, Department of Internal Medicine, University of Genoa; M. Bagnasco, MD, Medical and Radiometabolic Therapy Unit, Department of Internal Medicine, University of Genoa; A. Manfredi, MD, Unit of Internal Medicine and Immunology, IRCCS Ospedale San Raffaele; G.A. Ramirez, MD, Unit of Internal Medicine and Immunology, IRCCS Ospedale San Raffaele; P. Ramoino, MD, DISTAV, University of Genoa; P. Bianchini, MD, Department of Nanophysics, IIT; F. Puppo, MD, Department of Internal Medicine, University of Genoa; F. Pupo, MD, Department of Internal Medicine, University of Genoa; S. Negrini, MD, Department of Internal Medicine, University of Genoa; F. Mattana, MD, Division of Nephrology, University of Genoa and Policlinico San Martino; G. Emmi, MD, Lupus Clinic Department of Biomedicine, University of Florence, University Hospital Careggi; G. Garibotto, MD, Division of Nephrology, University of Genoa and Policlinico San Martino; D. Santoro, MD, Nephrology and Dialysis Unit, University of Messina and G. Martino Hospital; F. Scolari, MD, Division of Nephrology and Dialysis, University of Brescia and Ospedale di Montichiari; A. Ravelli, MD, Division of Paediatric Rheumatology, IRCCS Istituto Giannina Gaslini; A. Tincani, MD, Rheumatology and Clinical Immunology, Spedali Civili and University of Brescia; P. Cravedi, MD, Nephrology Division, Department of Medicine, Icahn School of Medicine at Mount Sinai; S. Volpi, MD, Division of Paediatric Rheumatology, IRCCS Istituto Giannina Gaslini; G. Candiano, PhD, Laboratory of Molecular Nephrology, IRCCS Istituto Giannina Gaslini; G.M. Ghiggeri, MD, PhD, Laboratory of Molecular Nephrology, and Division of Nephrology, Dialysis, and Transplantation, IRCCS Istituto Giannina Gaslini. GMarcoGhiggeri@ospedale-gaslini.ge.it.
Abstract
OBJECTIVE: Neutrophil extracellular traps (NET) expose modified antigens for autoantibodies in vasculitis. Little is known about levels and removal pathways of NET in systemic lupus erythematosus (SLE), especially in lupus nephritis (LN). We determined circulating levels and defined NET removal in large subsets of patients with incident SLE (iSLE), some of whom had new-onset nephritis. METHODS: Serum levels of NET (ELISA), DNase1/DNase1L3 (ELISA), and DNase activity (functional assay) were determined in 216 patients with iSLE [103 had incident LN (iLN)], in 50 patients with other primary glomerulonephritis, and in healthy controls. Ex vivo NET production by neutrophils purified from a random selection of patients was quantified as elastase/DNA release and by immunofluorescence techniques. RESULTS: Serum NET levels were very high in iSLE/iLN compared to all groups of controls and correlated with anti-dsDNA, C3-C4, and proteinuria; iLN had the highest levels. DNase activity was decreased in iLN compared to SLE (20% had one-half DNase activity) despite similar serum levels of DNase1/DNase1L3. In these cases, pretreatment of serum with protein A restored DNase efficiency; 1 patient was homozygous for a c.289_290delAC variant of DNASE1L3. Ex vivo NET production by neutrophils purified from LN, SLE, and normal controls was similar in all cases. CONCLUSION: Patients with iLN have increased circulating NET and reduced DNase activity, the latter being explained by the presence of inhibitory substances in circulation and/or by rare DNase1L3 mutations. Accumulation of NET derives from a multifactorial mechanism, and is associated and may contribute to disease severity in SLE, in particular to renal lesions. (Clinical trial registration: The Zeus study was registered at ClinicalTrials.gov, study number NCT02403115).
OBJECTIVE: Neutrophil extracellular traps (NET) expose modified antigens for autoantibodies in vasculitis. Little is known about levels and removal pathways of NET in systemic lupus erythematosus (SLE), especially in lupus nephritis (LN). We determined circulating levels and defined NET removal in large subsets of patients with incident SLE (iSLE), some of whom had new-onset nephritis. METHODS: Serum levels of NET (ELISA), DNase1/DNase1L3 (ELISA), and DNase activity (functional assay) were determined in 216 patients with iSLE [103 had incident LN (iLN)], in 50 patients with other primary glomerulonephritis, and in healthy controls. Ex vivo NET production by neutrophils purified from a random selection of patients was quantified as elastase/DNA release and by immunofluorescence techniques. RESULTS: Serum NET levels were very high in iSLE/iLN compared to all groups of controls and correlated with anti-dsDNA, C3-C4, and proteinuria; iLN had the highest levels. DNase activity was decreased in iLN compared to SLE (20% had one-half DNase activity) despite similar serum levels of DNase1/DNase1L3. In these cases, pretreatment of serum with protein A restored DNase efficiency; 1 patient was homozygous for a c.289_290delAC variant of DNASE1L3. Ex vivo NET production by neutrophils purified from LN, SLE, and normal controls was similar in all cases. CONCLUSION:Patients with iLN have increased circulating NET and reduced DNase activity, the latter being explained by the presence of inhibitory substances in circulation and/or by rare DNase1L3 mutations. Accumulation of NET derives from a multifactorial mechanism, and is associated and may contribute to disease severity in SLE, in particular to renal lesions. (Clinical trial registration: The Zeus study was registered at ClinicalTrials.gov, study number NCT02403115).
Authors: Abdul Hakkim; Barbara G Fürnrohr; Kerstin Amann; Britta Laube; Ulrike Abu Abed; Volker Brinkmann; Martin Herrmann; Reinhard E Voll; Arturo Zychlinsky Journal: Proc Natl Acad Sci U S A Date: 2010-05-03 Impact factor: 11.205
Authors: Stanley Moore; Hsin-Hsuan Juo; Christoffer T Nielsen; Helena Tyden; Anders A Bengtsson; Christian Lood Journal: J Rheumatol Date: 2019-12-15 Impact factor: 4.666
Authors: Maurizio Bruschi; Andrea Petretto; Laura Santucci; Augusto Vaglio; Federico Pratesi; Paola Migliorini; Roberta Bertelli; Chiara Lavarello; Martina Bartolucci; Giovanni Candiano; Marco Prunotto; Gian Marco Ghiggeri Journal: Sci Rep Date: 2019-05-28 Impact factor: 4.379
Authors: Gian Marco Ghiggeri; Matteo D'Alessandro; Domenico Bartolomeo; Maria Ludovica Degl'Innocenti; Alberto Magnasco; Francesca Lugani; Marco Prunotto; Maurizio Bruschi Journal: Int J Mol Sci Date: 2019-11-18 Impact factor: 5.923