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Morphological Characteristics of Rapid and Classical Netosis

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Abstract

The aim of this study was to determine the structural–morphological differences between rapid and classic netosis of human neutrophils. A method for studying the netosis process using lattices for transmission electron microscopy has been developed. Dynamics of blood neutrophil netosis and differences in the cell death processes induced by ionomycin and opsonized bacteria Staphylococcus aureus were studied with flow cytometry. Morphological features of rapid and classic netosis were examined with methods of scanning electron, fluorescence, and light microscopy. The structural and morphological similarities and differences between the two variants of netosis induced by opsonized S. aureus were described. It was revealed that classical netosis can induce the formation of extracellular traps in neighboring neutrophils. It was shown that the morphological features of extracellular traps can be used to differentially diagnose the development of rapid and classical netosis not only with high-resolution microscopy, but also with traditional light microscopy.

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ACKNOWLEDGMENTS

This study was partially performed on the equipment of the Centers for Collective Use New Materials and Resource-Saving Technologies and Center for Scanning Probe Microscopy. We are sincerely grateful to Prof. V.A. Vodeneev and Associated Professor I.V. Balalaeva for their help in fluorescence experiments.

Funding

This study was supported by the Russian Science Foundation, project no. 16-14-10179.

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Authors and Affiliations

  1. Research and Education Center for Physics of Solid-State Nanostructures of Lobachevsky Nizhny Novgorod National Research State University, 603950, Nizhny Novgorod, Russia

    S. N. Pleskova, E. N. Gorshkova, A. V. Boryakov & R. N. Kriukov

  2. Department of Nanotechnology and Biotechnology, Alekseev Nizhny Novgorod State Technical University, 603950, Nizhny Novgorod, Russia

    S. N. Pleskova

Authors
  1. S. N. Pleskova
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  2. E. N. Gorshkova
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  3. A. V. Boryakov
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  4. R. N. Kriukov
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Correspondence to S. N. Pleskova.

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Conflict of interests. The authors declare that they have no conflict of interest.

Statement of compliance with standards of research involving humans as subjects. All procedures involving human beings met the ethical standards of the Bioethics Commission of Lobachevsky Nizhny Novgorod National Research State University and the Helsinki Declaration of 1964 and its subsequent emendations. Informed voluntary consent was obtained from each of the participants.

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Translated by I. Fridlyanskaya

Abbreviations: AFM—atomic force microscopy, ROS—reactive oxygen species, BSA—bovine serum albumin, PBS—physiological buffered saline, MPO—myeloperoxidase, NG—neutrophil granulocyte, TEM—transmission electron microscopy, SEM—scanning electron microscopy, NET—neutrophil extracellular trap.

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Pleskova, S.N., Gorshkova, E.N., Boryakov, A.V. et al. Morphological Characteristics of Rapid and Classical Netosis. Cell Tiss. Biol. 14, 28–35 (2020). https://doi.org/10.1134/S1990519X20010083

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  • DOI: https://doi.org/10.1134/S1990519X20010083

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