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Abstract Number: 1774

Identification of Target Antigens of Anti-Endothelial Cell Antibodies in Patients with ANCA-Associated Systemic Vasculitis: A Proteomic Approach

Alexis Régent1, Hanadi Dib2, Guillaume Bussone1, Mathieu C. Tamby1, Nicolas Tamas2, Christian Federici3, Cédric Broussard3, Loic Guillevin4 and Luc Mouthon1, 1Internal Medicine, Hopital Cochin, Paris, France, 2Université Paris Descartes, Paris, France, 3Institut Cochin, INSERM U1016, CNRS UMR 8104, Université Paris Descartes, Paris, France, 4Department of Internal Medicine, Referral Center for Rare Autoimmune and Systemic Diseases, Hôpital Cochin, AP–HP, Université Paris Descartes, Paris, France, Paris, France

Meeting: 2012 ACR/ARHP Annual Meeting

Keywords: ANCA, autoantibodies, autoantigens and vasculitis, B cells

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Session Information

Session Title: B-cell Biology and Targets in Autoimmune Disease

Session Type: Abstract Submissions (ACR)

Background/Purpose: Anti-endothelial cell antibodies (AECA) are frequently detected in anti-neutrophil cytoplasm antibodies (ANCA)-associated systemic vasculitis (AAV) and are considered to play pathological roles but their antigenic specificities are still unknown. We used a proteomic approach combining two-dimensional electrophoresis and immunoblotting to identify the target antigens of AECA in patients with ANCA-associated vasculitis.

Methods: Sera from 30 ANCA-associated vasculitis patients (12 with Granulomatosis with polyangeitis (GPA), 9 with microscopic polyangeitis (MPA), 9 with Churg Strauss syndrome (CSS)), tested in pools of 3 sera, were compared to a sera pool from 12 healthy controls (HC). Serum IgG reactivity was analyzed by use of a 2-D electrophoresis and immunoblotting technique with normal human umbilical vein endothelial cell (HUVEC) antigens.

Results: Serum IgG in the HC sera pool recognized 85 protein spots and serum IgG from patients with AAV recognized 134±65 different protein spots. We focused on protein recognized by at least 3/4 pools of patients with GPA and 2/3 pools of patients with MPA and CSS and not by HC and identified 20, 7 and 8 proteins, respectively. In addition, among the 330 spots recognized by at least one pool of patients with AAV, ten different spots were recognized by at least 6/10 pools. Among identified proteins, IgG reactivity was detected against alpha-enolase, lamin A/C and protein disulfide-isomerase A3. Interestingly, Ingenuity Pathway Analysis revealed that most of these antigens interact with TGF-β, immunoglobins and inflammatory complexes such as Jun and MAPK

Conclusion: AECA detected in patients with AAV recognize cellular targets playing key roles in cell biology. Target antigens interact with protein and complexes known to play a crucial role in AAV pathophysiology.


Disclosure:

A. Régent,
None;

H. Dib,
None;

G. Bussone,
None;

M. C. Tamby,
None;

N. Tamas,
None;

C. Federici,
None;

C. Broussard,
None;

L. Guillevin,
None;

L. Mouthon,
None.

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