Antiaggregatory and proangiogenic effects of a novel recombinant human dual specificity anti-integrin antibody

Escher, R; Cung, T; Stutz, M; Haeberli, A; Djonov, V; Berchtold, P; Hlushchuk, R (2009). Antiaggregatory and proangiogenic effects of a novel recombinant human dual specificity anti-integrin antibody. Journal of thrombosis and haemostasis, 7(3), pp. 460-9. Oxford: Wiley-Blackwell 10.1111/j.1538-7836.2008.03251.x

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BACKGROUND: beta(3)-Integrins are involved in platelet aggregation via alpha(IIb)beta(3) [glycoprotein (GP)IIb-GPIIIa], and in angiogenesis via endothelial alpha(V)beta(3). Cross-reactive ligands with antiaggregatory and proangiogenic effects, both desirable in peripheral vasculopathies, have not yet been described. OBJECTIVES: In vitro and in vivo characterization of antiaggregatory and proangiogenic effects of two recombinant human Fab fragments, with emphasis on beta(3)-integrins. METHODS: Recombinant Fab fragments were obtained by phage display technology. Specificity, affinity and IC(50) were determined by immunodot assays, enzyme-linked immunosorbent assay (ELISA), and Scatchard plot analysis, and by means of human umbilical vein endothelial cells (HUVECs). Functional analyses included ELISA for interaction with fibrinogen binding to GPIIb-GPIIIa, flow cytometry for measurement of activation parameters and competitive inhibition experiments, human platelet aggregometry, and proliferation, tube formation and the chorioallantoic membrane (CAM) assay for measurement of angiogenic effects. RESULTS: We observed specific and high-affinity binding to an intact GPIIb-GPIIIa receptor complex of two human Fab autoantibody fragments, with no platelet activation. Dose-dependent fibrinogen binding to GPIIb-GPIIIa and platelet aggregation were completely inhibited. One Fab fragment was competitively inhibited by abciximab and its murine analog monoclonal antibody (mAb) 7E3, whereas the other Fab fragment bound to cultured HUVECs, suggesting cross-reactivity with alpha(V)beta(3), and also demonstrated proangiogenic effects in tube formation and CAM assays. CONCLUSIONS: These Fab fragments are the first entirely human anti-GPIIb-GPIIIa Fab fragments with full antiaggregatory properties; furthermore, they do not activate platelets. The unique dual-specificity anti-beta(3)-integrin Fab fragment may represent a new tool for the study and management of peripheral arterial vasculopathies.

Item Type:	Journal Article (Original Article)
Division/Institute:	?? DCD5A442B9C7E17DE0405C82790C4DE2 ??
UniBE Contributor:	Escher, Robert
ISSN:	1538-7836
ISBN:	19054322
Publisher:	Wiley-Blackwell
Language:	English
Submitter:	Factscience Import
Date Deposited:	04 Oct 2013 15:06
Last Modified:	05 Dec 2022 14:20
Publisher DOI:	10.1111/j.1538-7836.2008.03251.x
PubMed ID:	19054322
Web of Science ID:	000263256100012
URI:	https://boris.unibe.ch/id/eprint/28983 (FactScience: 132875)