Anti-ADAMTS13 Autoantibodies against Cryptic Epitopes in Immune-Mediated Thrombotic Thrombocytopenic Purpura

Elien Roose; Gestur Vidarsson; Kadri Kangro; Onno J. H. M. Verhagen; Ilaria Mancini; Linda Desender; Inge Pareyn; Nele Vandeputte; Aline Vandenbulcke; Chiara Vendramin; An-Sofie Schelpe; Jan Voorberg; Marie-Agnès Azerad; Laurent Gilardin; Marie Scully; Daan Dierickx; Hans Deckmyn; Simon F. De Meyer; Flora Peyvandi; Karen Vanhoorelbeke

doi:10.1055/s-0038-1669459

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2018; 118(10): 1729-1742
DOI: 10.1055/s-0038-1669459

Coagulation and Fibrinolysis

Georg Thieme Verlag KG Stuttgart · New York

Anti-ADAMTS13 Autoantibodies against Cryptic Epitopes in Immune-Mediated Thrombotic Thrombocytopenic Purpura

Elien Roose

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Gestur Vidarsson

²Department of Experimental Immunohematology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

,

Kadri Kangro

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Onno J. H. M. Verhagen

²Department of Experimental Immunohematology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

,

Ilaria Mancini

³Department of Pathophysiology and Transplantation, Università degli Studi di Milano, and Fondazione Luigi Villa, Milan, Italy

,

Linda Desender

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Inge Pareyn

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Nele Vandeputte

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Aline Vandenbulcke

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Chiara Vendramin

⁴Department of Haematology, University College London Hospital, London, United Kingdom

,

An-Sofie Schelpe

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Jan Voorberg

⁵Department of Plasma Proteins, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

,

Marie-Agnès Azerad

⁶Department of Hematology, CHU Brugmann, Brussels, Belgium

⁷Department of Hematology, CHU Liège, Liège, Belgium

,

Laurent Gilardin

⁸Département de Médicine Interne et Immunologie Clinique, Hôpital Pitié-Salpêtrière, Assistance Publique-Hôpitaux de Paris, Paris, France

,

Marie Scully

⁴Department of Haematology, University College London Hospital, London, United Kingdom

,

Daan Dierickx

⁹Department of Hematology, University Hospitals Leuven, Leuven, Belgium

,

Hans Deckmyn

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Simon F. De Meyer

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

,

Flora Peyvandi

³Department of Pathophysiology and Transplantation, Università degli Studi di Milano, and Fondazione Luigi Villa, Milan, Italy

¹⁰Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, Fondazione IRCCS Ca'Granda Ospedale Maggiore Policlinico, Milan, Italy

,

Karen Vanhoorelbeke

¹Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium

› Author Affiliations

Abstract

Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is characterized by severe ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type 1 repeats, member 13) deficiency, the presence of anti-ADAMTS13 autoantibodies and an open ADAMTS13 conformation with a cryptic epitope in the spacer domain exposed. A detailed knowledge of anti-ADAMTS13 autoantibodies will help identifying pathogenic antibodies and elucidating the cause of ADAMTS13 deficiency. We aimed at cloning anti-ADAMTS13 autoantibodies from iTTP patients to study their epitopes and inhibitory characteristics. We sorted anti-ADAMTS13 autoantibody expressing B cells from peripheral blood mononuclear cells of 13 iTTP patients to isolate anti-ADAMTS13 autoantibody sequences. Ninety-six B cell clones producing anti-ADAMTS13 autoantibodies were identified from which 30 immunoglobulin M (IgM) and 5 IgG sequences were obtained. For this study, we only cloned, expressed and purified the five IgG antibodies. In vitro characterization revealed that three of the five cloned IgG antibodies, TTP73–1, ELH2–1 and TR8C11, indeed recognize ADAMTS13. Epitope mapping showed that antibodies TTP73–1 and TR8C11 bind to the cysteine–spacer domains, while the antibody ELH2–1 recognizes the T2–T3 domains in ADAMTS13. None of the antibodies inhibited ADAMTS13 activity. Given the recent findings regarding the open ADAMTS13 conformation during acute iTTP, we studied if the cloned antibodies could recognize cryptic epitopes in ADAMTS13. Interestingly, all three antibodies recognize cryptic epitopes. In conclusion, we cloned three anti-ADAMTS13 autoantibodies from iTTP patients that recognize cryptic epitopes. Hence, these data nicely fit our recent finding that the conformation of ADAMTS13 is open during acute iTTP.

Keywords

thrombotic thrombocytopenic purpura - ADAMTS13 - anti-ADAMTS13 autoantibodies - cloning - cryptic epitopes

Full Text

References

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