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Clinical and Vaccine Immunology, May 2006, p. 575-583, Vol. 13, No. 5
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.5.575-583.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Changes in Activation States of Murine Polymorphonuclear Leukocytes (PMN) during Inflammation: a Comparison of Bone Marrow and Peritoneal Exudate PMN

Department of Preventive Veterinary Medicine and Animal Health, Nihon University School of Veterinary Medicine, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan,¹ Department of Rheumatology and Inflammation Research,² Department of Virology, Göteborg University, Guldhedsgatan 10, 413 46 Göteborg, Sweden³

Received 25 November 2005/ Returned for modification 24 January 2006/ Accepted 16 March 2006

To study different activation states in polymorphonuclear leukocytes (PMN) in mice, we compared the function of murine PMN obtained from the bone marrow (BMPMN) with those of PMN obtained by intraperitoneal induction with thioglycolate (TGPMN) or uric acid (UAPMN). When stimulated with chemotactic peptides, e.g., formyl-methionyl-leucyl-phenylalanine (fMLF), WKYMVM, or WKYMVm, the TGPMN and UAPMN showed greatly enhanced generation of reactive oxygen species (ROS) compared with BMPMN, which suggests that exudation to the peritoneum per se induces a primed state in the cells. The WKYMVm peptide was the most potent stimulant of ROS generation, and it desensitized for subsequent stimulation with fMLF or WKYMVM. This desensitization was broken by the addition of cytochalasin B. The TGPMN and UAPMN appeared to be fully primed, since no increase in response was induced by pretreatment with tumor necrosis factor (TNF-). In contrast, the BMPMN response was increased 2.5- to 3-fold. The differences in oxidative responses were supported by degranulation studies. Preincubation with TNF- promoted CR3 expression on BMPMN, and this level of expression was also enhanced by WKYMVm. In contrast, CR3 expression on untreated TGPMN and UAPMN was already similar to that on TNF--primed BMPMN and could be only slightly enhanced by TNF- treatment. Taken together, these results indicate that BMPMN are in a resting state and have the capacity to become primed, while peritoneal exudate PMN are already fully primed upon isolation. These results have major implications for murine neutrophil research and show the importance of defining which PMN subsets to use when investigating murine models.