Azoximer bromide effect on the neutrophil extracellular traps formation
B.V. Pinegin¹, Yu.A. Dagil², N.V. Vorobieva³, M.V. Pashchenkov<sup>1

1</sup>National Research Center — Institute of Immunology Federal Medical-Biological Agency of Russia, Moscow
² LLC “Petrovax Pharm”, Moscow
³Lomonosov Moscow State University

In 2004, a new immunological process, consisting of DNA strands release framed with cationic proteins and peptides during neutrophil activation, was presented. These structures were called neutrophil extracellular traps (NETs). Along with the presence of NETs positive properties, it was shown that NETs have a serious negative effect associated with their protein composition, consisting of 70% of positively charged histones and 20–30% of cationic proteins such as α-defensins and peptide LL-37 (which in the same manner as histones have a toxic effect on human tissue outside the cellular space) in relation to the infection protection. Currently, it is proved that NETs are involved in numerous pathological conditions, including those with respiratory infections, significantly aggravating the course of the disease.
Aim: to study the effect of azoximer bromide (under «Polyoxidonium» trade name) on the NETs formation in vitro.
Patients and Methods: neutrophils of healthy donors were isolated by centrifugation in one-step Ficoll-Hypaque density gradient. Polyoxidonium in 10, 100, 500, or 1000 μg/ml dosages was added to neutrophils 1 hour before the phorbol myristate acetate (PMA) introduction into the wells. Fluorescence microscopy was used to detect NETs. The preparations were stained with Syber Green.
Results: a new effect of the Polyoxidonium drug has been identified. It consisted in reducing the activated neutrophils ability to secrete macromolecular complexes including DNA, granular, nuclear and cytoplasmic proteins, and antimicrobial proteins as well, which are equally toxic for both prokaryotic and eukaryotic cells. A decrease in the NETs formation and release has made a significant contribution to the therapeutic and prophylactic effect of Polyoxidonium. Also, it explained its high clinical efficacy in the treatment and prevention of acute and chronic respiratory viral infections along with the immunomodulating and detoxifying properties.
Conclusion: conducted researches revealed fundamentally new therapeutic properties of Polyoxidonium, namely, its ability to reduce the NETs formation by activated neutrophils with full preservation of its immunostimulating and detoxifying properties.

Keywords: neutrophil extracellular traps, NETosis, respiratory diseases, ARVI, human neutrophils, azoximer bromide, Polyoxidonium.
For citation: Pinegin B.V., Dagil Yu.A., Vorobieva N.V., Pashchenkov M.V. Azoximer bromide effect on the neutrophil extracellular traps formation. RMJ. 2019;1(II):42–46.