Inhibition of thrombin-induced platelet aggregation using human single-chain Fv antibodies specific for TREM-like transcript-1

Barbara Giomarelli; Valance A. Washington; Maia M. Chisholm; Laura Quigley; James B. McMahon; Toshiyuki Mori; Daniel W. McVicar

doi:10.1160/TH06-08-0456

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2007; 97(06): 955-963
DOI: 10.1160/TH06-08-0456

Platelets and Blood Cells

Schattauer GmbH

Inhibition of thrombin-induced platelet aggregation using human single-chain Fv antibodies specific for TREM-like transcript-1

Barbara Giomarelli^*

¹Molecular Targets Development Program

,

Valance A. Washington^*

²Cancer and Inflammation Program– Center for Cancer Research, National Cancer Institute, NCI-Frederick, Frederick, Maryland, USA

³Department of Anatomy and Cell Biology, Universidad Central del Caribe, Bayamón, Puerto Rico

,

Maia M. Chisholm

²Cancer and Inflammation Program– Center for Cancer Research, National Cancer Institute, NCI-Frederick, Frederick, Maryland, USA

,

Laura Quigley

²Cancer and Inflammation Program– Center for Cancer Research, National Cancer Institute, NCI-Frederick, Frederick, Maryland, USA

,

James B. McMahon

¹Molecular Targets Development Program

,

Toshiyuki Mori^#

¹Molecular Targets Development Program

,

Daniel W. McVicar

²Cancer and Inflammation Program– Center for Cancer Research, National Cancer Institute, NCI-Frederick, Frederick, Maryland, USA

› Author Affiliations

Abstract

Summary

TREM-like transcript-1 (TLT-1) is a novel platelet membrane receptor, which has been recently characterized in mice. TLT-1 is expressed exclusively in platelets and megakaryocytes, and its expression is dramatically upregulated upon platelet activation, suggesting that it plays a unique role in hemostasis and/or thrombosis. In this study we identified and characterized highly specific human monoclonal antibodies that bind to TLT-1 by screening a naïve library of single chain Fv fragments (scFvs) displayed on filamentous phage (Thomlinson I library). These scFvs detected plate-bound TLT-1, captured soluble TLT-1, and readily reacted with cell-bound TLT-1 on transfectants and primary human platelets. Most importantly, anti-TLT-1 scFvs inhibited thrombin-mediated human platelet aggregation. This inhibition was specific for thrombin-induced aggregation and was reversible with higher doses of agonist. These data are the first to demonstrate a biological role for TLT-1 and its potential as a therapeutic target. The human scFvs isolated in this study may represent novel anti-platelet therapeutic agents.

Keywords

Triggering receptors expressed on myeloid cells (TREM)-like transcript-1 (TLT-1) - anti-thrombosis therapy - human antibodies - single chain fragments (scFvs) - phage display

Full Text

References

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