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Chelation of Free Zn2+ Impairs Chemotaxis, Phagocytosis, Oxidative Burst, Degranulation, and Cytokine Production by Neutrophil Granulocytes

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Abstract

Neutrophil granulocytes are the largest leukocyte population in the blood and major players in the innate immune response. Impaired neutrophil function has been reported in in vivo studies with zinc-deficient human subjects and experimental animals. Moreover, in vitro formation of neutrophil extracellular traps has been shown to depend on free intracellular Zn2+. This study investigates the requirement of Zn2+ for several other essential neutrophil functions, such as chemotaxis, phagocytosis, cytokine production, and degranulation. To exclude artifacts resulting from indirect effects of zinc deprivation, such as impaired hematopoietic development and influences of other immune cells, direct effects of zinc deprivation were tested in vitro using cells isolated from healthy human donors. Chelation of Zn2+ by the membrane permeable chelator N,N,N′,N′-tetrakis-(2-pyridylmethyl)-ethylenediamine (TPEN) reduced granulocyte migration toward N-formyl-l-methionyl-l-leucyl-l-phenylalanine (fMLF) and IL-8, indicating a role of free intracellular Zn2+ in chemotaxis. However, a direct action of Zn2+ as a chemoattractant, as previously reported by others, was not observed. Similar to chemotaxis, phagocytosis, oxidative burst, and granule release were also impaired in TPEN-treated granulocytes. Moreover, Zn2+ contributes to the regulatory role of neutrophil granulocytes in the inflammatory response by affecting the cytokine production by these cells. TPEN inhibited the lipopolysaccharide-induced secretion of chemotactic IL-8 and also anti-inflammatory IL-1ra. In conclusion, free intracellular Zn2+ plays essential roles in multiple neutrophil functions, affecting extravasation to the site of the infection, uptake and killing of microorganisms, and inflammation.

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Acknowledgments

We thank Dr. Jörg Häseler, TU Berlin, for critically reading the manuscript. This study was supported in part by German Research Council (DFG) grant HA4318/3 to H. H. Furthermore, R. H. received a scholarship from the German Academic Exchange Service (DAAD).

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Authors and Affiliations

  1. Institute of Immunology, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany

    Rafah Hasan, Lothar Rink & Hajo Haase

  2. Department of Food Chemistry and Toxicology, Berlin Institute of Technology, Gustav-Meyer-Allee 25, 13355, Berlin, Germany

    Hajo Haase

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  1. Rafah Hasan
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Correspondence to Hajo Haase.

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Hasan, R., Rink, L. & Haase, H. Chelation of Free Zn2+ Impairs Chemotaxis, Phagocytosis, Oxidative Burst, Degranulation, and Cytokine Production by Neutrophil Granulocytes. Biol Trace Elem Res 171, 79–88 (2016). https://doi.org/10.1007/s12011-015-0515-0

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