Influence of iron- and zinc-chelating agents
on neutrophil extracellular trap formation

Weronika Kuźmicka; Aneta Moskalik; Aneta Manda-Handzlik; Urszula Demkow; Małgorzata Wachowska; Olga Ciepiela

doi:10.5114/ceji.2021.106985

eISSN: 1644-4124
ISSN: 1426-3912

Central European Journal of Immunology

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2/2021
vol. 46

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abstract:

Experimental immunology

Influence of iron- and zinc-chelating agents on neutrophil extracellular trap formation

Weronika Kuźmicka

^{1, 2}

,

Aneta Moskalik

²

,

Aneta Manda-Handzlik

¹

,

Urszula Demkow

¹

,

Małgorzata Wachowska

¹

,

Olga Ciepiela

³

Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Warsaw, Poland
Postgraduate School of Molecular Medicine, Medical University of Warsaw, Warsaw, Poland
Department of Laboratory Medicine, Medical University of Warsaw, Warsaw, Poland

Cent Eur J Immunol 2021; 46 (2): 135-139

DOI: https://doi.org/10.5114/ceji.2021.106985

Online publish date: 2021/06/18

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Release of neutrophil extracellular traps (NETs) is one of the neutrophils’ mechanisms involved in the response to infection. NETs are released from the cell in response to a biological or synthetic stimulus to entrap, immobilize and kill pathogens. Metal ions and metal binding proteins were identified in the structure of NETs, but their role in NET release remains unclear. The aim of this study was to assess how lack of iron and zinc generated by ion sequestration using chelators affects NET release. Neutrophils were isolated from whole blood or buffy coats of healthy blood donors by density gradient centrifugation and incubated with zinc chelators: 20 µM N,N,N,N-Tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), 40 µM diethylenetriaminepentaacetic acid (DTPA) or iron chelators: 400 µM deferoxamine mesylate salt (DFO) and 50 µM iminodiacetic acid (IDA). Next, 100 nM phorbol 12-myristate 13-acetate (PMA) was added to stimulate release of NETs. The amount of released DNA was measured by fluorometry and NETs were visualized by immunofluorescence microscopy. This study demonstrates that iron and zinc chelators are able to modulate NET release. Here we show that preincubation of neutrophils with TPEN and IDA inhibits NET release in cells stimulated with PMA. On the other hand, DFO stimulates NET release. Incubation of cells with DTPA does not affect release of NETs.

keywords:

Influence of iron- and zinc-chelating agents on neutrophil extracellular trap formation

Weronika Kuźmicka 1, 2 , Aneta Moskalik 2 , Aneta Manda-Handzlik 1 , Urszula Demkow 1 , Małgorzata Wachowska 1 , Olga Ciepiela 3

deferoxamine, iminodiacetic acid, DTPA, TPEN, neutrophil extracellular traps

Weronika Kuźmicka

^{1, 2}

,

Aneta Moskalik

²

,

Aneta Manda-Handzlik

¹

,

Urszula Demkow

¹

,

Małgorzata Wachowska

¹

,

Olga Ciepiela

³