Role of Antioxidants in Horse Serum-mediated Vasculitis in Swine: Potential Relevance to Early Treatment in Mitigation of Coronary Arteritis in Kawasaki Disease

Background: Kawasaki Disease (KD) is the leading cause of acquired heart disease in the US. We have demonstrated the critical role of innate immune responses via IL-1R/MyD88 signaling in the Lactobacillus casei cell wall extract (LCWE)-induced KD mouse model. The diversity and composition of microflora (both bacterial and fungal) have been associated with the regulation and alterations of immune responses and various pathologies. However, the role of gut microbiota in immunopathology of KD has not been investigated. Objective: To evaluate the role of gut microflora in development of coronary arteritis, and vascular abnormalities in KD mouse model. Methods and Results: We investigated the role of gut microflora in the LCWE-induced KD mouse model, using Specific-Pathogen Free (SPF) and Germ Free (GF) mice (C57BL/6). GF mice showed a significant decrease of KD lesions, including coronary arteritis compared with SPF mice. The development of LCWE-induced AAA, which we recently discovered in this mouse model, was also markedly diminished in GF mice. In addition to GF mice, we also investigated the specific role of commensal fungi, and determined whether altered fungal burden in this KD mouse model contributes to disease severity. To deplete fungi in the gut microflora, we exposed pregnant SPF mice and their offspring to fluconazole (antifungal) in their drinking water for 5 wks and induced KD. The fluconazole treated mice had significantly reduced coronary arteritis, and AAA compared to controls. Since Dectin-1 has emerged as a key receptor that recognizes β-1,3-glucans found in the cell wall of nearly all fungi, we next induced KD in Dectin-1 deficient mice. Dectin-1 deficient mice also had significantly reduced KD lesions such as coronary arteritis compared with WT mice. Conclusions: We demonstrate here that gut microflora play a critical role in the development of KD vasculitis in LCWE-induced mouse model. Our results suggest that fungi in the intestinal microbiota may specifically control the induction and severity of KD vasculitis, which may be mediated by Dectin-1. These findings provide a new perspective on the potential role of the microbiome in KD pathogenesis and may offer new diagnostic and therapeutic strategies for KD patients.

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Abstract 65: T cell Differentiation in Human Kawasaki Disease and a Murine Model of Coronary Arteritis

Circulation ◽

10.1161/circ.131.suppl_2.65 ◽

2015 ◽

Vol 131 (suppl_2) ◽

Author(s):

I-Chun Lin ◽

Hsuan-Chang Kuo ◽

Ying-Jui Lin ◽

Shao-Ju Chien ◽

Chien-Fu Huang ◽

...

Keyword(s):

Cell Differentiation ◽

T Cell ◽

Kawasaki Disease ◽

Murine Model ◽

Th17 Cells ◽

T Cell Differentiation ◽

Ivig Treatment ◽

Th1 Cells ◽

Coronary Arteritis

The role of T cell differentiation in the immunopathogenesis of Kawasaki disease (KD) remains unclear. The aim of this study is to elucidate the role of T cell subsets in coronary artery lesion (CAL) of KD. Peripheral blood was obtained in 10 patients with acute KD before and 1 week after intravenous gamma-immunoglobulin (IVIG) treatment and in 20 patients with past history of KD for more than 5~20 years. Meanwhile, induction of coronary arteritis was performed on wild type BALB/c mice by Lactobacillus casei cell wall extract (LCWE). Human peripheral blood leukocytes were analyzed by using flow cytometric analysis and murine hearts were examined for immunofluorescence study and for RNA expression levels. Results: Compared to the febrile controls, KD patients prior to IVIG treatment had increased percentage of CD3 + /CD4 + /interferon-γ + (T helper 1, Th1) cells and CD3 + /CD4 + /interleukin-17A + (Th17) cells (mean ± SD, 1.36% ± 1.39% and 0.51% ± 0.25%, respectively) among Th cells (CD3 + /CD4 + ). Both increases declined after IVIG treatment (0.71% ± 0.74% and 0.33% ± 0.18%) despite no statistically difference by Mann-Whitney test. None of these 10 acute KD patients developed CAL after IVIG treatment. However, patients with previous KD and definite CALs (n= 11) seemed to have higher percentage of Th17 cells (0.50% ± .025% versus 0.35% ± 0.23%) but similar level of Th1 cells (0.93% ± 0.51% versus 1.05% ± 0.64%) when compared to those without CAL (n= 9). Murine cardiac tissues displayed the presence of Th1 (double-stained with CD3 and T-bet) and Th17 cells (double-stained with CD3 and RORγt) during days 7 and 14 after LCWE treatment but not in PBS-treated mice. Compatible with these, cardiac mRNA levels showed both increased levels of IFN-γ and IL17A mRNA in LCWE-treated mice. Conclusions: Our initial data suggests that specific T cell differentiation into Th1 and Th17 cells occurred in both human KD and mice stimulated with LCWE. IVIG treatment was associated with the recovery of such T cell differentiation. However, the clinical application to predict IVIG responsiveness and future CAL development by such increase in the peripheral Th17 remains unclear. Further studies to elucidate the detailed immune regulation of these T subsets on CAL are warrant by using this LCWE murine model.

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Abstract 71: Abdominal Aorta Dilatation and Aneurysm in Kawasaki Disease Vasculitis Mouse Model: Role of IL-1 Signaling

Circulation ◽

10.1161/circ.131.suppl_2.71 ◽

2015 ◽

Vol 131 (suppl_2) ◽

Author(s):

Daiko Wakita ◽

Youngho Lee ◽

Kenichi Shimada ◽

Shuang Chen ◽

Timothy R Crother ◽

...

Keyword(s):

Coronary Artery ◽

Kawasaki Disease ◽

Mouse Model ◽

Abdominal Aorta ◽

Vascular Complications ◽

Aortic Dilatation ◽

Coronary Arteritis ◽

Abdominal Aortic ◽

Aorta Dilatation

Background: Kawasaki disease (KD) is the most common cause of acute systemic vasculitis and acquired cardiac disease among US children. KD causes coronary artery aneurysms in up to 25% of untreated patients, and less frequently aneurysms in other systemic arteries including the abdominal aorta. Lactobacillus casei cell wall extract (LCWE)-induced KD mouse model mimics histopathologically the coronary artery lesions seen in KD patients. Objective: To evaluate the development of abdominal aorta dilatation and aneurysm in KD mouse model and investigate the role of IL-1 signaling. Methods and Results: We investigated the incidence and progression of abdominal aorta aneurysm (AAA) and dilatation in the KD model at 1, 2, 5 weeks. Over 80% of the mice developed significant dilation of abdominal aorta at 2 wks with progressively greater dilatation at 5 wks, with greater severity in males. KD mice showed fusiform and saccular AAA, which were always below the renal artery. Immunohistochemistry showed significant intimal proliferation, massive myofibroblastic proliferation that breaks the elastin layer, infiltration of large numbers of neutrophils and macrophages into the media and adventitia. IL-1R- or IL-1beta-deficient mice were completely protected from the KD associated abdominal aorta dilatation and AAA. IL-1R antagonist (Anakinra) significantly prevented the abdominal aorta dilatation and AAA (in addition to blocking coronary arteritis) in the KD mice. Conclusions: We report a new model of AAA and aortic dilatation in the LCWE-induced KD mouse model. These studies suggest that in children with KD the incidence of abdominal aortic dilatation and AAA maybe higher than currently appreciated, thus requiring prospective studies to determine the frequency of these vascular complications. Our findings also demonstrate that IL-1 plays an important role in development of LCWE-induced abdominal aortic lesions and blockade of IL-1 signaling may be a promising therapeutic target not only for KD vasculitis and coronary arteritis, but also for abdominal aorta dilatation and AAA associated with the disease.

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The role of anti-endothelial cell antibodies in Kawasaki disease -in vitro and in vivo studies

Clinical & Experimental Immunology ◽

10.1046/j.1365-2249.2002.02000.x ◽

2002 ◽

Vol 130 (2) ◽

pp. 233-240 ◽

Cited By ~ 50

Author(s):

E. GRUNEBAUM ◽

M. BLANK ◽

S. COHEN ◽

A. AFEK ◽

J. KOPOLOVIC ◽

...

Keyword(s):

Endothelial Cell ◽

Kawasaki Disease ◽

In Vivo Studies

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Early anti IL-1 treatment replaces steroids in refractory Kawasaki disease: clinical experience from two case reports

Therapeutic Advances in Musculoskeletal Disease ◽

10.1177/1759720x211002593 ◽

2021 ◽

Vol 13 ◽

pp. 1759720X2110025

Author(s):

Maria Vincenza Mastrolia ◽

Giulia Abbati ◽

Claudia Signorino ◽

Ilaria Maccora ◽

Edoardo Marrani ◽

...

Keyword(s):

Kawasaki Disease ◽

Clinical Evidence ◽

Case Reports ◽

Therapeutic Option ◽

Intravenous Immunoglobulins ◽

Second Line Therapy ◽

Coronary Artery Aneurysms ◽

Coronary Arteritis ◽

Line Therapy ◽

Prior Use

Refractory Kawasaki disease (KD) is related to a major risk of coronary arteries abnormalities and its treatment is not standardized. In this regard, anakinra (ANA), an interleukin (IL)-1 receptor antagonist, represents an emerging therapeutic option. We report two cases of children, diagnosed with KD, nonresponsive to two doses of intravenous immunoglobulins, successfully treated with ANA, without a prior use of steroids. Patient 2 developed a coronary dilatation, that improved significantly after ANA therapy. Our experience highlights IL-1 blockade effectiveness in reducing KD inflammation and suggests ANA adoption as second-line therapy, with a timesaving and steroid-sparing strategy. Our results, combined with the evidence of the IL-1 key role in KD and coronary arteritis pathogenesis and to the recent clinical evidence reported by the KAWAKINRA trial, encourage an earlier recourse to ANA in patients with refractory KD, in order to fight inflammation, and to treat and prevent the development of coronary artery aneurysms. Further studies are needed to better define the place of IL-1 blockade in KD step-up treatment.

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Abstract 636: Gut Microflora Influences Pathology in the Kawasaki Disease (KD) Vasculitis Mouse Model

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.35.suppl_1.636 ◽

2015 ◽

Vol 35 (suppl_1) ◽

Author(s):

Daiko Wakita ◽

Yosuke Kurashima ◽

Yoshihiro Takasato ◽

Youngho Lee ◽

Kenichi Shimada ◽

...

Keyword(s):

Mouse Model ◽

Immune Responses ◽

Critical Role ◽

Gut Microflora ◽

Vascular Abnormalities ◽

Coronary Arteritis ◽

Specific Pathogen ◽

New Perspective ◽

Bacteria And Fungi

Background: KD is the leading cause of acquired heart disease in the US. We have demonstrated the critical role of innate immune responses via IL-1R/MyD88 signaling in the Lactobacillus casei cell wall extract (LCWE)-induced KD mouse model. The diversity and composition of microflora (both bacterial and fungal) have been associated with the regulation and alterations of immune responses and various pathologies. However, the role of gut microbiota in immunopathology of KD has not been investigated. Objective: To evaluate the role of gut microflora in development of coronary arteritis, and vascular abnormalities in KD mouse model. Methods and Results: We investigated the role of gut microflora in the LCWE-induced KD mouse model, using Specific-Pathogen Free (SPF) and Germ Free (GF) mice (C57BL/6). GF mice showed a significant decrease of KD lesions, including coronary arteritis compared with SPF mice. The development of LCWE-induced AAA, which we recently discovered in this mouse model, was also markedly diminished in GF mice. In addition to GF mice, we also investigated the specific role of commensal bacteria and/or fungi, and determined whether altered microorganism burden in this KD mouse model contributes to disease severity. To deplete bacteria and/or fungi in the gut microflora, we exposed pregnant SPF mice and their offspring to antibiotics cocktail (Abx) or antifungal drug (fluconazole; Fluc) in their drinking water for 5 wks and induced KD. The mice treated with Abx or Fluc had significantly reduced coronary arteritis and AAA compared to controls. The Abx plus Fluc administration showed marked decrease of KD vasculitis. Conclusions: We demonstrate here that gut microflora play a critical role in the development of KD vasculitis in LCWE-induced mouse model. Our results suggest that both bacteria and fungi in the intestinal microbiota may control the induction and severity of KD vasculitis. These findings provide a new perspective on the potential role of the microbiome in KD pathogenesis and may offer new diagnostic and therapeutic strategies for KD patients.

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