HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Published online before print December 14, 2006

This Article Full Text (Reprint (PDF)) All Versions of this Article: jlb.0806513v1 81/3/672    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Stie, J. Articles by Jesaitis, A. J. Search for Related Content PubMed PubMed Citation Articles by Stie, J. Articles by Jesaitis, A. J.

© by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb.0806513

Received for publication August 10, 2006.

Accepted for publication November 17, 2006.

Article

Reorganization of the human neutrophil plasma membrane is associated with functional priming: implications for neutrophil preparations

Montana State University, Department of Microbiology, Bozeman, Montana, USA

^@ To whom correspondence should be addressed. E-mail: umbaj{at}montana.edu.

	Abstract

Changes in the functional and plasma membrane organizational states of human neutrophils were examined using two isolation procedures, which may simulate altered physiological states in vivo. A gelatin-based method of blood-neutrophil isolation was used to model in vivo priming, and neutrophils isolated by this method were compared with control populations prepared by a pyrogen-free, dextran-based method. Gelatin-prepared neutrophils were functionally primed for adherence and agonist-stimulated superoxide generation relative to unprimed, control neutrophils. The organizational state of the membrane cortex was examined by mapping the subcellular distribution of select cortical and transmembrane proteins by several methods, including subcellular fractionation, indirect immunofluorescence, and compositional analysis of Triton X-100-insoluble membrane skeleton preparations. Filamentous actin, fodrin, and the fodrin anchor CD45 were largely cytoplasmic in unprimed neutrophils but translocated to plasma membranes upon priming, whereas CD43 and ezrin were exclusively surface-associated in both populations. Isopycnic sucrose density gradient analysis of N₂-cavitated neutrophils revealed a major shift in the distribution of surface-associated transmembrane and membrane cortical components relative to the plasma membrane marker alkaline phosphatase in primed but not unprimed neutrophils. Similar results were obtained after neutrophil stimulation with known priming agents, LPS, TNF-, or GM-CSF. Together, these results may suggest that priming of suspended, circulating neutrophils is associated with a large-scale reorganization of the plasma membrane and associated membrane cortex in a process that is independent of cellular adhesion and gross morphologic polarization.

Key Words: inflammation • leukocyte • granulocyte • degranulation • cortical cytoskeleton

This article has been cited by other articles:

				S. M. Uriarte, K. R. McLeish, and R. A. Ward Anti-proteinase 3 antibodies both stimulate and prime human neutrophils Nephrol. Dial. Transplant., October 24, 2008; (2008) gfn580v1. [Abstract] [Full Text] [PDF]

				M. Riesselman, H. M. Miettinen, J. M. Gripentrog, C. I. Lord, B. Mumey, E. A. Dratz, J. Stie, R. M. Taylor, and A. J. Jesaitis C-Terminal Tail Phosphorylation of N-Formyl Peptide Receptor: Differential Recognition of Two Neutrophil Chemoattractant Receptors by Monoclonal Antibodies NFPR1 and NFPR2 J. Immunol., August 15, 2007; 179(4): 2520 - 2531. [Abstract] [Full Text] [PDF]