SEARCH:   Advanced

Blood, 15 April 2007, Vol. 109, No. 8, pp. 3521-3528. Prepublished online as a Blood First Edition Paper on December 12, 2006; DOI 10.1182/blood-2006-03-007179. This Article Full Text Full Text (PDF) Supplemental Table and Figures Supplemental Videos All Versions of this Article: blood-2006-03-007179v1 109/8/3521    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Eisenhardt, S. U. Articles by Peter, K. Search for Related Content PubMed PubMed Citation Articles by Eisenhardt, S. U. Articles by Peter, K. Related Collections Cell Adhesion and Motility Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  PHAGOCYTES Generation of activation-specific human anti-Mß2 single-chain antibodies as potential diagnostic tools and therapeutic agents Steffen U. Eisenhardt1, Meike Schwarz2, Nils Schallner2, Juliana Soosairajah1, Nicole Bassler1, Dexing Huang1, Christoph Bode2, and Karlheinz Peter1 1 Baker Heart Research Institute, Melbourne, Victoria, Australia; 2 Department of Cardiology, University of Freiburg, Germany The leukocyte integrin Mac-1 (Mß2) plays a pivotal role in inflammation and host defense. Upon leukocyte activation, Mac-1 undergoes a conformational change exposing interaction sites for multiple ligands. We aimed to generate single-chain antibodies (scFv's) directed against activation-specific Mac-1 ligand-binding sites. Using human scFv phage libraries, we developed subtractive strategies with depletion of phages binding to nonactivated Mac-1 and selection of phages binding to activated Mac-1, using monocytes as well as CHO cells transfected with native or mutated, activated Mac-1. Three scFv clones demonstrated exclusive binding to activated Mac-1. Mac-1 binding of the ligands fibrinogen, heparin, and ICAM-1, but not C3bi, was inhibited. Using alanine substitutions, the paratope was identified within the heavy chain HCDR3s of the scFv's. The epitope was localized to Lys245-Arg261 of the M I-domain. In a pilot study with septicemic patients, we provide initial support for the use of these scFv's as markers of monocyte activation and as potential diagnostic tools. Potential therapeutic use was tested in adhesion assays under static and flow conditions demonstrating the selective blockade of activated monocytes only. Furthermore, scFv HCDR3–derived peptides retain selectivity for the activated integrin, providing a unique template for the potential development of inhibitors that are specific for the activated Mac-1. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Blood orchestrates a leukocyte integrin trio Joel S. Bennett Blood 2007 109: 3137-3138. [Full Text] [PDF] This article has been cited by other articles: C. von zur Muhlen, D. von Elverfeldt, J.A. Moeller, R.P. Choudhury, D. Paul, C.E. Hagemeyer, M. Olschewski, A. Becker, I. Neudorfer, N. Bassler, et al. Magnetic Resonance Imaging Contrast Agent Targeted Toward Activated Platelets Allows In Vivo Detection of Thrombosis and Monitoring of Thrombolysis Circulation, July 15, 2008; 118(3): 258 - 267. [Abstract] [Full Text] [PDF] I. Ahrens, C. Ellwanger, B. K. Smith, N. Bassler, Y. C. Chen, I. Neudorfer, A. Ludwig, C. Bode, and K. Peter Selenium supplementation induces metalloproteinase-dependent L-selectin shedding from monocytes J. Leukoc. Biol., June 1, 2008; 83(6): 1388 - 1395. [Abstract] [Full Text] [PDF]

	Copyright © 2007 by American Society of Hematology         Online ISSN: 1528-0020