Open Access Research article

Autoantibodies to angiotensin and endothelin receptors in systemic sclerosis induce cellular and systemic events associated with disease pathogenesis

Angela Kill12, Christoph Tabeling3, Reinmar Undeutsch12, Anja A Kühl4, Jeannine Günther12, Mislav Radic25, Mike O Becker2, Harald Heidecke6, Margitta Worm7, Martin Witzenrath3, Gerd-Rüdiger Burmester2, Duska Dragun8 and Gabriela Riemekasten12*

Author Affiliations

1 German Rheumatism Research Centre (DRFZ), A Leibniz Institute, Berlin, Germany

2 Department of Rheumatology and Clinical Immunology, University Hospital Charité, Luisenstraße 13, Berlin 10117, Germany

3 Department of Infectious Diseases and Pulmonary Medicine, University Hospital Charité, Berlin, Germany

4 Department of Inner Medicine, University Hospital Charité, Berlin, Germany

5 Department of Rheumatology and Clinical Immunology, University Hospital Split, Split, Croatia

6 CellTrend GmbH, Luckenwalde, Germany

7 Department of Dermatology, University Hospital Charité, Berlin, Germany

8 Department of Nephrology and Intensive Care Medicine, University Hospital Charité, Berlin, Germany

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Arthritis Research & Therapy 2014, 16:R29  doi:10.1186/ar4457

Published: 28 January 2014

Abstract

Introduction

Vasculopathy, inflammatory fibrosis and functional autoantibodies (Abs) are major manifestations of systemic sclerosis (SSc). Abs directed against the angiotensin II type 1 receptor (AT1R) and endothelin-1 type A receptor (ETAR) are associated with characteristic disease features including vascular, inflammatory, and fibrotic complications indicating their role in SSc pathogenesis. Therefore, the impact of anti-AT1R and anti-ETAR Abs on initiation of inflammation and fibrosis was analyzed.

Methods

Anti-AT1R and anti-ETAR Ab-positive immunoglobulin G (IgG) from SSc patients (SSc-IgG) was used for experiments. Healthy donor IgG served as a normal control, and AT1R and ETAR activation was inhibited by antagonists. Protein expression was measured with ELISA, mRNA expression with real time-PCR, endothelial repair with a scratch assay, and collagen expression with immunocytochemistry. Transendothelial neutrophil migration was measured with a culture insert system, and neutrophil ROS activation with immunofluorescence. Neutrophils in bronchoalveolar lavage fluids (BALFs) were analyzed microscopically after passive transfer of SSc-IgG or NC-IgG into naïve C57BL/6J mice. KC plasma levels were quantified by a suspension array system. Histologic analyses were performed by using light microscopy.

Results

Anti-AT1R and anti-ETAR Ab-positive SSc-IgG induced activation of human microvascular endothelial cells (HMEC-1). Elevated protein and mRNA levels of the proinflammatory chemokine interleukin-8 (IL-8, CXCL8) and elevated mRNA levels of the vascular cell adhesion molecule-1 (VCAM-1) were induced in HMEC-1. Furthermore, activation of HMEC-1 with SSc-IgG increased neutrophil migration through an endothelial cell layer and activation of reactive oxygen species (ROS). SSc-IgG decreased HMEC-1 wound repair and induced type I collagen production in healthy donor skin fibroblasts. Effects of migration, wound repair, and collagen expression were dependent on the Ab-levels. Passive transfer of anti-AT1R and anti-ETAR Ab-positive SSc-IgG into naïve C57BL/6J mice increased neutrophil BALF counts. In parallel, increased levels of the murine functional IL-8 homologue, chemokine KC, were found in the plasma of SSc-IgG-treated mice as well as structural alterations of the lungs.

Conclusions

We conclude that angiotensin and endothelin-receptor activation via anti-AT1R and anti-ETAR Abs mediate pathogenic effects, indicating their contribution to pathogenesis of SSc. Therefore, anti-AT1R and anti-ETAR Abs could provide novel targets for therapeutic intervention in the treatment of SSc.