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Thromb Haemost 2011; 106(05): 947-958
DOI: 10.1160/TH11-05-0337
New Technologies, Diagnostic Tools and Drugs
Schattauer GmbH

Novel recombinant glycosylphosphatidylinositol (GPI)-anchored ADAMTS13 and variants for assessment of anti-ADAMTS13 autoantibodies in patients with thrombotic thrombocytopenic purpura

Dengju Li
Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia and The University of Pennsylvania Medical Center, Philadelphia, Pennsylvania, USA
,
Juan Xiao
Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia and The University of Pennsylvania Medical Center, Philadelphia, Pennsylvania, USA
,
Michele Paessler
Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia and The University of Pennsylvania Medical Center, Philadelphia, Pennsylvania, USA
,
X. Long Zheng
Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia and The University of Pennsylvania Medical Center, Philadelphia, Pennsylvania, USA
› Author Affiliations Financial support: This study is supported in part by grants from National Institute of Health (HL074124) and American Heart Association-Established Investigator Award. DL is supported in part by a fellowship from Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Further Information

Publication History

Received: 17 May 2011

Accepted after major revision: 19 July 2011

Publication Date:
23 November 2017 (online)

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Summary

Immunoglobulin Gs (IgGs) against ADAMTS13 are major causes of acquired (idiopathic) thrombotic thrombocytopenic purpura (TTP). We report here a novel cell-based assay using glycosylphosphatidylinositol (GPI)-anchored ADAMTS13 or variants expressed on cell membrane for assessment of autoantibodies in patients with TTP. We showed that IgGs from all 26 patients with acquired TTP bound to cells expressing a GPI anchored full-length ADAMTS13 (gFL) and a variant truncated after the spacer domain (gS). Also, IgGs from 25/26 (96.7%) of these TTP patients bound to cells expressing a GPI-anchored C-terminal fragment, TSP1 2–8 plus CUB (gT2C). In contrast, none of the 20 healthy blood donors showed detectable binding of their IgGs to the cells expressing gFL, gS, and gT2C. A moderate, but statistically significant correlation was observed between plasma concentrations of anti-ADAMTS13 IgG and positive cells expressing gFL (r=0.65), gS (r=0.67), and gT2C (r=0.42). These results suggest that the microtiter-plate assay and the cell-based assay may detect differential antigenic epitopes. Moreover, antigens clustered on cell membranes may enhance antibody binding affinity, thereby increasing analytical sensitivity. Finally, our assay was able to determine kinetic changes of plasma levels of anti-ADAMTS13 IgGs in TTP patients during plasma therapy. Together, our findings suggest that the novel cell-based assay may be applicable for rapid identification and mapping of anti-ADAMTS13 autoantibodies in patients with acquired TTP.

Keywords

von Willebrand factor cleaving protease - thrombotic microangio-pathies - diagnostic test - autoantibody

 

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