Document Type : Original Article

Authors

Wuxi Children’s Hospital Affiliated, Jiangnan University, China.

Abstract

Background: Kawasaki disease (KD) is a vasculitis associated with vascular injury and autoimmune response. Inflammatory factors stimulate neutrophils to produce web-like structures called neutrophil extracellular traps (NETs). Citrullinated histone 3 (H3Cit) is one of the main protein components of neutrophil extracellular traps involved in the process of NETosis. The levels of NETs and H3Cit in the KD are not known.
Objective: To determine the changes in the levels of NETs and H3Cit in KD.
Methods: Children with KD were recruited and divided into the acute KD and the sub-acute KD group according to the disease phase and whether intravenous immunoglobulin (IVIG) was used or not. Peripheral venous blood was taken before and after the IVIG administration and sent for the examination of NETs by flow cytometry. The level of H3Cit was measured by enzyme-linked immunosorbent assay (ELISA).
Results: The counts of NETs in the acute KD group significantly increased compared with the healthy controls (p<0.01). The level of H3Cit was significantly higher in the acute KD group than in the healthy control subjects. Of note, both the counts of NETs and the level of H3Cit decreased in the KD patients treated with IVIG compared with the acute KD group (p<0.01).
Conclusion: Acute KD is characterized by an increased formation of NETs and high levels of H3Cit. IVIG significantly inhibited NETs formation and also reduced the level of plasma H3Cit in children with KD.

Keywords

  1. 1.Kawasaki T, Singh S.Kawasaki disease-the journey over 50 years:1967-2017.Int J Rheum Dis. 2018;21:7-9.

    1. Rowley AH, Shulman ST.The Epidemiology and Pathogenesis of Kawasaki Disease.Front Pediatr. 2018;6:374.

    3.Viner, R. M. & Whittaker, E.Kawasaki-like disease: emerging complication during the COVID-19 pandemic.Lancet.2020;395:1741-1743.

    1. Jiang, L., Tang, K.,Levin,M.& Irfan.OCOVID-19 and multisystem inflammatory syndrome in children and adolescents published correction appears in Lancet Infect Dis.Lancet Infect Dis.2020; 20:e276-e288.
    2. Kitamura, S. & Tsuda, E. Significance of Coronary Revascularization for Coronary-Artery Obstructive Lesions Due to Kawasaki Disease.Children (Basel).2019;6:16.
    3. Orenstein JM, Shulman ST, Fox LM, Baker SC, Takahashi M, Bhatti TR, Russo PA, Mierau GW,et al.Three linked vasculopathic processes characterize Kawasaki disease: a light and transmission electron microscopic study.2012;PLoS One7:e38998.
    4. Mostafavi, N., Haghjooy-Javanmard, S., Presidend, N., Manssori, N.S. & Kelishadi,R.Persistence of endothelial cell damage late after Kawasaki disease in patients without coronary artery complications. Adv. Biomed. Res. 2015;4:25.
    5. 8. Newburger JW, Takahashi M, Gerber MA, Gewitz MH, Tani LY, Burns JC. et a.Diagnosis, treatment, and long-term management of Kawasaki disease: a statement for health professionals from the Committee on Rheumatic Fever, Endocarditis, and Kawasaki Disease, Council on Cardiovascular Disease in the Young, American Heart Association. 2004; 114:1708-33.
    6. Takahashi, K., Wakayama, M., Yokouchi, Y., Wakaya, M. & Youkouchi, Y. Neutrophilic involvement in the damage to coronary arteries in acute stage of Kawasaki disease. Pediatr Int. 2005;47:305-310.
    7. Tsujimoto H, Takeshita S, Nakatani K, Kawamura Y, Tokutomi T, Sekine I.Delayed apoptosis of circulating neutrophils in Kawasaki disease.Clin.Exp Immuno.2001;l126:355-364.
    8. Hu J, Qian W, Yu Z, Xu T, Ju L, Hua Q,et al.Increased Neutrophil Respiratory urst predicts the Risk of Coronary Artery Lesion in Kawasaki Disease.Front Pediatr. 2020;8:391.
    9. Brinkmann V, Reichard U, Goosmann C, Fauler, B., Uhlemann, Y., Weiss, D. S.et al.Neutrophil extracellular traps kill bacteria. Science .2004;303:1532-1535.
    10. Grayson, P.C. & Kaplan, M.J.At the Bench: Neutrophil extracellular traps (NETs) highlight novel aspects of innate immune system involvement in autoimmune diseases.J Leukoc Biol.2016;99:253-264.
    11. Hofbauer,T.M.,Mangold,A.,Scherz,T.,Seidl,V.,Panzenböck,A.,Ondracek.Neutrophil extracellular traps and fibrocytes in ST-segment elevation myocardial infarction.Basic Res Cardiol. 2019;114:33.

    15.Ducroux,C.,Di Meglio, L.,Loyau,S.,Delbosc,S.,Boisseau,W.,Deschildre,C.et al.Thrombus Neutrophil Extracellular Traps Content Impair tPA-Induced Thrombolysis in Acute Ischemic Stroke. Stroke.2018;49:754-757.

    1. Kapoor S, Opneja A, Nayak L.The role of neutrophils in thrombosis.Thromb Res.2018;170:87-96.

    17.Thålin, C.,Daleskog,M.,Göransson,S.P.,Schatzberg,D.,Lasselin,J.,Laska,A.et al.Validation of an enzyme-linked immunosorbent assay for the quantification of citrullinated histone H3 as a marker for neutrophil extracellular traps in human plasma.Immunol Res. 2017;65:706-712.

    18.Deng,Q.,Pan,B.,Alam,H.B.,Liang,Y.,Wu,Z.,Liu,B et al.Citrullinated Histone H3 as a Therapeutic Target for Endotoxic Shock in Mice.Front Immunol.2020;10:2957.

    19.Paues Göranson,S.,Thålin,C.,Lundström,A.,Hållström,L.,Lasselin,J.,Wallén,H.,et al.Circulating H3Cit is elevated in a human model of endotoxemia and can be detected bound to microvesicles.Sci Rep. 2018;8:12641.

    20.Da Silva, R. F., Baptista, D.,Roth, A.,Miteva,K.,Burger,F.,Vuilleumier,N.et al.Anti-Apolipoprotein A-1 IgG Influences Neutrophil Extracellular Trap Content at Distinct Regions of Human Carotid Plaques. Int. J. Mol Sci. 2020;21:7721.

    21.Gavillet, M., Martinod, K., Renella, R., Harris, C., Shapiro, N. I., Wagner, D.,et al.Flow cytometric assay for direct quantification of neutrophil extracellular traps in blood samples. Am J Hematol. 2015;90:1155-1158.

    22.Fousert E,Toes R,Desai J.Neutrophil extracellular traps (NETs) take the central stage in driving autoimmune responsesv.Cells.2020;9:915.

    23.Saffarzadeh,M.,Juenemann,C.,Queisser, M.A.,Lochnit,G.,Barreto,G.,Galuska,S.P, et al.Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones.Plos one. 2012;7:e32366.

    24.Distelmaier K, Winter MP, Dragschitz F, et al.Prognostic value of culprit site neutrophils in acute coronary syndrome. Eur J Clin Invest.2014;44:257-265.

    25.Stark,K.,Philippi,V.,Stockhausen,S.,Busse,J.,Antonelli,A.,Miller,M.,et al.Disulfide HMGB1 derived from platelets coordinates venous thrombosis in mice. Blood.2016;128:2435-2449.

    26.Delabranche,X.,Stiel,L.,Severac,F.,Galoisy,A.C.,Mauvieux,L.,Zobairi,F.,et al.Evidence of NETosis in septic shock induced disseminated intravascular coagulation.Shock. 2017; 47:313-317.

    27.Yoshida Y, Takeshita S, Kawamura Y, Kanai T, Tsujita Y, Nonoyama S.Enhanced formation of neutrophil extracellular traps in Kawasaki disease.Pediatr Res. 2020;87:998-1004.

    28.Chao,C.H.,Wu,W.C.,Lai,Y.C.,Tsai,P.J.,Perng,G.C.,Lin,Y.S.,et al.Dengue virus nonstructural protein 1 activates platelets via Toll-like receptor 4, leading to thrombocytopenia and hemorrhage. PLoS Pathog.2019;15:e1007625.

    29.Kaneko K, Takahashi M, Yoshimura K et al. (2012) Intra- venous immunoglobulin counteracts oxidative stress in Kawasaki disease. Pediatr Cardiol 33 (7), 1086–8.

    30.Ballow, M.The IgG molecule as a biological immune response modifier: mechanisms of action of intravenous immune serum globulin in autoimmune and inflammatory disorders.J Allergy Clin Immunol. 2011;127:315-23.

    31.Okubo,K.,Kamiya, M.,Urano,Y.,Nishi,H.,Herter,J.M., Mayadas,T.,et al.Lactoferrin suppresses neutrophil extracellular traps release in inflammation. EBioMedicine. 2016;10:204-215.

    32.Shida,H.,Nakazawa, D.,Tateyama,Y.,Miyoshi,A.,Kusunoki,Y.,Hattanda,F.,et al.The presence of anti-lactoferrin antibodies in a subgroup of eosinophilic granulomatosis with polyangiitis patients and their possible contribution to enhancement of neutrophil extracellular trap formation.Front Immunol. 2016;7: 636.

    33.Silk, E.,Zhao,H.,Weng,H.&Ma,D.The role of extracellular histone in organ injury.Cell Death Dis. 2017;8:e2812.

    34.Xu,J.,Zhang,X.,Pelayo,R.,Monestier,M.,Ammollo,C.T.,Semeraro,F.,et al.Extracellular histones are major mediators of death in sepsis.Nat Med.2009;15:1318-1321.

    35.Parackova Z,Zentsova I,Malcova H,Cebecauerova D,Sediva A,Horvath R.Increased histone citrullination in juvenile idiopathic arthritis. Front Med (Lausanne).2022;19:971121.