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J. Biol. Chem., Vol. 282, Issue 32, 23603-23612, August 10, 2007

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The Neutrophil-specific Antigen CD177 Is a Counter-receptor for Platelet Endothelial Cell Adhesion Molecule-1 (CD31)^*

Ulrich J. H. Sachs¹, Cornelia L. Andrei-Selmer, Amudhan Maniar, Timo Weiss, Cathy Paddock, Valeria V. Orlova^¶, Eun Young Choi^¶, Peter J. Newman, Klaus T. Preissner^||, Triantafyllos Chavakis^¶, and Sentot Santoso²

From the Institute for Clinical Immunology and Transfusion Medicine and the ^||Institute for Biochemistry, Justus Liebig University, Langhansstrasse 7, Giessen D-35392, Germany, the Blood Research Institute, Blood Center of Wisconsin, Milwaukee, Wisconsin 53233, and the ^¶Experimental Immunology Branch, NCI, National Institutes of Health, Bethesda, Maryland 20892

Human neutrophil-specific CD177 (NB1 and PRV-1) has been reported to be up-regulated in a number of inflammatory settings, including bacterial infection and granulocyte-colony-stimulating factor application. Little is known about its function. By flow cytometry and immunoprecipitation studies, we identified platelet endothelial cell adhesion molecule-1 (PECAM-1) as a binding partner of CD177. Real-time protein-protein analysis using surface plasmon resonance confirmed a cation-dependent, specific interaction between CD177 and the heterophilic domains of PECAM-1. Monoclonal antibodies against CD177 and against PECAM-1 domain 6 inhibited adhesion of U937 cells stably expressing CD177 to immobilized PECAM-1. Transendothelial migration of human neutrophils was also inhibited by these antibodies. Our findings provide direct evidence that neutrophil-specific CD177 is a heterophilic binding partner of PECAM-1. This interaction may constitute a new pathway that participates in neutrophil transmigration.

Received for publication, February 6, 2007 , and in revised form, May 31, 2007.

^* This work was supported in part by the Deutsche Forschungsgemeinschaft (Grant SFB 547) and by an intramural research program, NCI, National Institutes of Health (to T. C.). Parts of this work contribute to the doctoral theses of Amudhan Maniar and Timo Weiss. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ A fellow of the German Foundation for Haemotherapeutic Research (Bonn, Germany).

² To whom correspondence should be addressed. Tel.: 49-641-994-1518; Fax: 49-641-994-1529; E-mail: Sentot.santoso{at}med.uni-giessen.de.

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