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Inflammation. 2001 Feb;25(1):53-9.

Decreased leukotriene release from neutrophils after severe trauma: role of immature cells.

Author information

1
Department of Surgery, Surgical Research, BG Kliniken Bergmannsheil, Bochum, Germany.

Abstract

Polymorphonuclear granulocytes (PMN) play a key role in host defense against microbial infections. After severe trauma PMN show cellular dysfunctions including chemotactic migration, phagocytosis, and bacterial killing. In these settings the contribution of the cellular maturation stage compared to functional activities has not been investigated. Polymorphonuclear granulocytes are potent producers of lipid mediators via the 5-lipoxygenase (5-LO) pathway (leukotrienes, LTs) which exert important proinflammatory and immunoregulatory activities. We analyzed leukotriene generation from PMN-fractions (N = 23) of 15 polytrauma patients in comparison to 17 healthy donor cell fractions and correlated this lipid mediator release to the hematopoietic maturation stage of respective PMN. Polymorphonuclear granulocytes were isolated from EDTA-anticoagulated peripheral blood employing a one step procedure based on a discontinuous double Ficoll-gradient. Cells (5 x 10(6)/500 microl phosphate-buffered saline) were stimulated for 20 min at 37 degrees C with 1 microM Ca-ionophor A23187 in the presence of 1 mM Ca++ and 0.5 mM Mg++. Leukotrienes were analyzed by reversed-phase HPLC. Expression of 5-lipoxygenase (5-LO) was additionally determined by Western blot. Maturation stage of PMN was quantitated by Pappenheim-staining of cell smears. After polytrauma the generation of leukotrienes from PMN was individually diminished. Synthesis of enzymatically formed metabolites (LTB4, OH-LTB4 and COOH-LTB4) was concomitantly reduced. The decresaed leukotriene synthesis strongly correlated (r2 = 0.907, P < 0.0001) to the occurrence of immature PMN (mostly band cells). The expression of 5-lipoxygenase in PMN fractions consisting mainly of band cells was decreased. Our results provide evidence that posttraumatic granulocyte dysfunction is partly due to immature functional cell capacities.

PMID:
11293666
DOI:
10.1023/a:1007027712387
[Indexed for MEDLINE]

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