title: Kawasaki disease-specific molecules in the sera are linked to microbe-associated molecular patterns in the biofilms.
abstract: BACKGROUND
  Kawasaki disease (KD) is a systemic vasculitis of unknown etiology. The innate immune system is involved in its pathophysiology at the acute phase. We have recently established a novel murine model of KD coronary arteritis by oral administration of a synthetic microbe-associated molecular pattern (MAMP). On the hypothesis that specific MAMPs exist in KD sera, we have searched them to identify KD-specific molecules and to assess the pathogenesis.
  
  
  METHODS
  We performed liquid chromatography-mass spectrometry (LC-MS) analysis of fractionated serum samples from 117 patients with KD and 106 controls. Microbiological and LC-MS evaluation of biofilm samples were also performed.
  
  
  RESULTS
  KD samples elicited proinflammatory cytokine responses from human coronary artery endothelial cells (HCAECs). By LC-MS analysis of KD serum samples collected at 3 different periods, we detected a variety of KD-specific molecules in the lipophilic fractions that showed distinct m/z and MS/MS fragmentation patterns in each cluster. Serum KD-specific molecules showed m/z and MS/MS fragmentation patterns almost identical to those of MAMPs obtained from the biofilms formed in vitro (common MAMPs from Bacillus cereus, Yersinia pseudotuberculosis and Staphylococcus aureus) at the 1st study period, and from the biofilms formed in vivo (common MAMPs from Bacillus cereus, Bacillus subtilis/Bacillus cereus/Yersinia pseudotuberculosis and Staphylococcus aureus) at the 2nd and 3rd periods. The biofilm extracts from Bacillus cereus, Bacillus subtilis, Yersinia pseudotuberculosis and Staphylococcus aureus also induced proinflammatory cytokines by HCAECs. By the experiments with IgG affinity chromatography, some of these serum KD-specific molecules bound to IgG.
  
  
  CONCLUSIONS
  We herein conclude that serum KD-specific molecules were mostly derived from biofilms and possessed molecular structures common to MAMPs from Bacillus cereus, Bacillus subtilis, Yersinia pseudotuberculosis and Staphylococcus aureus. Discovery of these KD-specific molecules might offer novel insight into the diagnosis and management of KD as well as its pathogenesis.
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kusuda
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Nakashima
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Murata
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Kanno
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Hisanori
authors.initials: H
authors.lastname: Nishio
authors.affiliation: Department of Bacteriology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Mitsumasa
authors.initials: M
authors.lastname: Saito
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Tamami
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Yamamura
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Sakai
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Takada
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Miyamoto
authors.affiliation: Department of Pediatric Infectious Disease, Fukuoka Children's Hospital and Medical Center for Infectious Disease, Fukuoka, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Mizuno
authors.affiliation: Department of Pediatrics, Kawasaki Medical School Hospital, Okayama, Japan.
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Ouchi
authors.affiliation: Department of Pediatrics, Kurashiki Central Hospital, Okayama, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Waki
authors.affiliation: Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Toshiro
authors.initials: T
authors.lastname: Hara
mesh_terms: D001409:Bacillus cereus / Q000502:physiology; D001412:Bacillus subtilis / Q000502:physiology; D018441:Biofilms; D015415:Biomarkers / Q000097:blood*; D016022:Case-Control Studies; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid / Q000379:methods; D016207:Cytokines / Q000302:isolation & purification; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D009080:Mucocutaneous Lymph Node Syndrome / Q000097:blood* / Q000276:immunology / Q000382:microbiology* / Q000473:pathology; D013211:Staphylococcus aureus / Q000502:physiology; D015011:Yersinia pseudotuberculosis / Q000502:physiology
chemical_list: D015415:Biomarkers; D016207:Cytokines