title: Chelation of Free Zn²⁺ Impairs Chemotaxis, Phagocytosis, Oxidative Burst, Degranulation, and Cytokine Production by Neutrophil Granulocytes.
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 Zn(2+). This study investigates the requirement of Zn(2+) 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 Zn(2+) 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 Zn(2+) in chemotaxis. However, a direct action of Zn(2+) 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, Zn(2+) 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 Zn(2+) plays essential roles in multiple neutrophil functions, affecting extravasation to the site of the infection, uptake and killing of microorganisms, and inflammation.
authors.affiliation: Institute of Immunology, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany.
authors.forename: Rafah
authors.initials: R
authors.lastname: Hasan
authors.affiliation: Institute of Immunology, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany.
authors.forename: Lothar
authors.initials: L
authors.lastname: Rink
authors.affiliation: Institute of Immunology, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany. haase@TU-Berlin.de.|Department of Food Chemistry and Toxicology, Berlin Institute of Technology, Gustav-Meyer-Allee 25, 13355, Berlin, Germany. haase@TU-Berlin.de.
authors.forename: Hajo
authors.initials: H
authors.lastname: Haase
mesh_terms: D015550:Cell Degranulation; D002614:Chelating Agents / Q000737:chemistry / Q000378:metabolism*; D002633:Chemotaxis; D016207:Cytokines / Q000096:biosynthesis*; D006801:Humans; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism*; D010587:Phagocytosis; D016897:Respiratory Burst; D015032:Zinc / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002614:Chelating Agents; D016207:Cytokines; D015032:Zinc
keywords: Chemotaxis; Neutrophil granulocytes; Oxidative burst; Phagocytosis; TPEN; Zinc