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Published online before print November 7, 2005
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* Institute for Surgical Research and
Medical Policlinic-Innenstadt, Arbeitsgruppe Klinische Biochemie, University of Munich, Germany
1 Correspondence: Institute for Surgical Research, University of Munich, Marchioninistr. 27, D-81377 Munich, Germany. E-mail: andrej.khandoga{at}med.uni-muenchen.de
Leukocyte infiltration of reperfused tissue is a key event in the pathogenesis of ischemia-reperfusion. However, the role of chemokine receptors Ccr1, Ccr2, and Ccr5 for each single step of the postischemic recruitment process of leukocytes has not yet been characterized. Leukocyte rolling, firm adherence, transendothelial, and extravascular migration were analyzed in the cremaster muscle of anaesthetized C57BL/6 mice using near-infrared reflected light oblique transillumination microscopy. Prior to 30 min of ischemia as well as at 5, 30, 60, 90, and 120 min after onset of reperfusion, migration parameters were determined in wild-type, Ccr1–/–, Ccr2–/–, and Ccr5–/– mice. Sham-operated wild-type mice without ischemia were used as controls. No differences were detected in numbers of rolling leukocytes among groups. In contrast, the number of firmly adherent leukocytes was increased significantly in wild-type mice as compared with sham-operated mice throughout the entire reperfusion phase. Already after 5 min of reperfusion, this increase was reduced significantly in Ccr1–/– and Ccr5–/– mice, whereas only in Ccr2–/– mice, was adherence attenuated significantly at 120 min after onset of reperfusion. Furthermore, after 120 min of reperfusion, the number of transmigrated leukocytes (>80% Ly-6G+ neutrophils) was elevated in wild-type mice as compared with sham-operated animals. This elevation was significantly lower in Ccr1–/–, Ccr2–/–, and Ccr5–/– mice. Leukocyte extravascular migration distances were comparable among groups. In conclusion, these in vivo data demonstrate that Ccr1, Ccr2, and Ccr5 mediate the postischemic recruitment of neutrophils through effects on intravascular adherence and subsequent transmigration.
Key Words: ischemia-reperfusion • leukocytes • transmigration • MIP-1
• MCP-1 • RANTES
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