Qi You1, Dong Mei He1, Guo Fang Shu2, Bo Cao3, Yong Quan Xia4, Yun Xing1, Min Ni5, Ji Fang Chen6, Shu Li Shi7, Harvest F Gu8, Yu Liu9, Jie Wu1. 1. Minigene Pharmacy Laboratory, School of Life Science and Technology, China Pharmaceutical University, Nanjing, China. 2. Clinical Laboratory, Zhongda Hospital Affiliated to Southeast University, Nanjing, China. 3. Clinical Laboratory, Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China. 4. Clinical Laboratory, Gulou Hospital Affiliated to Medical College of Nanjing University, Nanjing, China. 5. Department of Endocrinology and Rheumatology, Nanjing Tongren Hospital Affiliated to Southeast University, Nanjing, China. 6. Department of Endocrinology, Jurong People's Hospital, Zhenjiang, China. 7. Clinical Laboratory, Jurong People's Hospital, Zhenjiang, China. 8. Center for Pathophysiology, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China. 9. Department of Endocrinology and Metabolism, Sir Run Run Hospital Affiliated to Nanjing Medical University, Nanjing, China.
Abstract
BACKGROUND: The aim of this study was to investigate the association of the formation of neutrophil extracellular traps (NETs) with gut leakage in type 1 (T1D) and type 2 diabetes (T2D). METHODS: In all, 105 subjects (56 T1D, 49 T2D) were included in the study. Eight biomarkers of NET formation and gut leakage (ie, protein arginine deiminase type 4 [PAD4], neutrophil elastase [NE], proteinase 3 [PR3], complement 5a [C5a], α1 -antitrypsin [AAT], DNase I, zonulin, and lipopolysaccharide [LPS]) were measured in serum samples by ELISA. Neutrophils were isolated and stimulated by phorbol myristate acetate to form NETs in vitro. Neutrophil intracellular contents were then collected and used as antigens to detect anti-neutrophil cytoplasmic antibodies (ANCA) in the serum. RESULTS: There was an increase in NET-associated proteins (PAD4, NE, PR3, C5a, AAT and DNase I) in new-onset T1D patients but not in those with T2D. Of PAD4, NE, and PR3, PAD4 was found to be the most sensitive biomarker for the diagnosis of T1D. Furthermore, circulating levels of zonulin and LPS were not only increased, but were also strongly correlated with NET formation and ANCA generation in T1D patients. CONCLUSIONS: This study provides evidence that increased formation of NETs, particularly PAD4, is closely associated with gut leakage in T1D but not T2D, and suggests that microorganisms and the release of neutrophil cytoplasmic antigen during the formation of NETs may be involved in the pathogenesis of T1D.
BACKGROUND: The aim of this study was to investigate the association of the formation of neutrophil extracellular traps (NETs) with gut leakage in type 1 (T1D) and type 2 diabetes (T2D). METHODS: In all, 105 subjects (56 T1D, 49 T2D) were included in the study. Eight biomarkers of NET formation and gut leakage (ie, protein arginine deiminase type 4 [PAD4], neutrophil elastase [NE], proteinase 3 [PR3], complement 5a [C5a], α1 -antitrypsin [AAT], DNase I, zonulin, and lipopolysaccharide [LPS]) were measured in serum samples by ELISA. Neutrophils were isolated and stimulated by phorbol myristate acetate to form NETs in vitro. Neutrophil intracellular contents were then collected and used as antigens to detect anti-neutrophil cytoplasmic antibodies (ANCA) in the serum. RESULTS: There was an increase in NET-associated proteins (PAD4, NE, PR3, C5a, AAT and DNase I) in new-onset T1D patients but not in those with T2D. Of PAD4, NE, and PR3, PAD4 was found to be the most sensitive biomarker for the diagnosis of T1D. Furthermore, circulating levels of zonulin and LPS were not only increased, but were also strongly correlated with NET formation and ANCA generation in T1D patients. CONCLUSIONS: This study provides evidence that increased formation of NETs, particularly PAD4, is closely associated with gut leakage in T1D but not T2D, and suggests that microorganisms and the release of neutrophil cytoplasmic antigen during the formation of NETs may be involved in the pathogenesis of T1D.
Authors: Jonas Schulte-Schrepping; Nico Reusch; Daniela Paclik; Kevin Baßler; Stephan Schlickeiser; Bowen Zhang; Benjamin Krämer; Tobias Krammer; Sophia Brumhard; Lorenzo Bonaguro; Elena De Domenico; Daniel Wendisch; Martin Grasshoff; Theodore S Kapellos; Michael Beckstette; Tal Pecht; Adem Saglam; Oliver Dietrich; Henrik E Mei; Axel R Schulz; Claudia Conrad; Désirée Kunkel; Ehsan Vafadarnejad; Cheng-Jian Xu; Arik Horne; Miriam Herbert; Anna Drews; Charlotte Thibeault; Moritz Pfeiffer; Stefan Hippenstiel; Andreas Hocke; Holger Müller-Redetzky; Katrin-Moira Heim; Felix Machleidt; Alexander Uhrig; Laure Bosquillon de Jarcy; Linda Jürgens; Miriam Stegemann; Christoph R Glösenkamp; Hans-Dieter Volk; Christine Goffinet; Markus Landthaler; Emanuel Wyler; Philipp Georg; Maria Schneider; Chantip Dang-Heine; Nick Neuwinger; Kai Kappert; Rudolf Tauber; Victor Corman; Jan Raabe; Kim Melanie Kaiser; Michael To Vinh; Gereon Rieke; Christian Meisel; Thomas Ulas; Matthias Becker; Robert Geffers; Martin Witzenrath; Christian Drosten; Norbert Suttorp; Christof von Kalle; Florian Kurth; Kristian Händler; Joachim L Schultze; Anna C Aschenbrenner; Yang Li; Jacob Nattermann; Birgit Sawitzki; Antoine-Emmanuel Saliba; Leif Erik Sander Journal: Cell Date: 2020-08-05 Impact factor: 41.582