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Neutrophil surface phenotype and oxygen metabolism alteration during priming


I.V. Obraztsov - Research Scientist, Department of Microbiology and Immunology Research, A.N. Ryzhikh State Scientific Centre for Coloproctology (Moscow)
M.A. Sukhina - Head of Department, Microbiology and Immunology Research, A.N. Ryzhikh State Scientific Centre for Coloproctology (Moscow)
S.A. Frolov - Deputy Director, A.N. Ryzhikh State Scientific Centre for Coloproctology (Moscow)
M.I. Rogovets - Post-graduate Student, N.I. Pirogov Russian National Research Medical University (Moscow)
A.Yu. Ryabov - Ph.D. (Phys.-Math.), Research Scientist, Department of Physics, Ludwig-Maximilians-Universität München (Germany)
E.V. Proskurnina - Ph.D. (Chem.), Associate Professor, Department of Medical Biophysics, Faculty of Fundamental Medicine, Lomonosov Moscow State University

Neutrophils have many important functions, such as phagocytosis, production of oxygen reactive species, the release of cytokines, and the formation of extracellular networks. The implementation of these functions is possible due to priming. The aim of this work was to evaluate the dynamics of the surface phenotype and oxidative metabolism alterations during neutrophil priming.
Neutrophils of the whole blood from ten healthy individuals were primed with phorbol-12-myristate-13 acetate (PMA) for 0–25 minutes (with the step of 5 minutes); N-formyl-methionyl-leucyl-phenylalanine (fMLP) was used to induce the oxygen burst. Oxidative metabolism was studied by the kinetic chemiluminescent (CL) assay (luminol as a CL enhancer), the sur-face phenotype was assessed by flow cytometry. We determined the levels of expression of immunoglobulin Fc receptors (FcγRIII, CD16 and FcγRI, CD64) and adhesion molecules (L-selectin, CD62L, αM-integrin, CD11b, Lewis X, CD15).
Neutrophil response to priming occurs within 5–10 minutes. The initially homogeneous pool of neutrophils contains sub-populations that react to priming at different rates and at different levels. We revealed the decrease of CD62L expression and the increase of CD15 and CD11b expression. The expression of CD64 and CD16 does not change. We determined in-tensification of oxygen metabolism, which is accompanied by an increase in the proportion of extracellular reactive oxygen species production.
To sum, the significant heterogeneity and extreme lability of the human neutrophil population allows foreseeing the pros-pects of using the functional characteristics of these cells as biomarkers of various inflammatory-septic complications in clinical practice.

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