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.
- Vorob'eva N.V. NADPH oksidaza nejtrofilov i zabolevaniya, svyazannye s eyo disfunkciej // Immunologiya. 2013. T. 34. № 4. S. 232–238.
- Vorob'eva N.V., Pinegin B.V. Nejtrofil'nye vnekletochnye lovushki: mekhanizmy obrazovaniya, rol' v norme i pri patologii (obzor) // Biohimiya. 2014. T. 79. № 12. S. 1580–1591.
- Bae M.H., Park S.H., Park C.J. et al. Flow cytometric measurement of respiratory burst activity and surface expression of neutrophils for septic patient prognosis // Cytometry Part B: Clinical Cytometry. 2016. V. 90. № 4. P. 368–375.
- Coates T.D. Neutrophils под ред. R.M. Kliegman, Stanton, B. F., St Geme, J.W. et al. Philadelphia, PA 19103-2899: Elsevier Inc. 2016. 1032–1035 s.
- El-Benna J., Hurtado-Nedelec M., Marzaioli V., Marie J. C. et al. Priming of the neutrophil respiratory burst: role in host defense and inflammation // Immunological Reviews. 2016. V. 273. № 1. P. 180–193.
- Genestet C., Le Gouellec A., Chaker H. et al. Scavenging of reactive oxygen species by tryptophan metabolites helps Pseudomonas aeruginosa escape neutrophil killing // Free Radical Biology and Medicine. 2014. V. 73. P. 400–410.
- Hong C.-W. Current Understanding in Neutrophil Differentiation and Heterogeneity // Immune Network. 2017. V. 17. № 5. P. 298.
- Hsu K.-H., Chan M. C., Wang J. M. et al. Comparison of Fcγ receptor expression on neutrophils with procalcitonin for the diagnosis of sepsis in critically ill patients // Respirology. 2011. V. 16. № 1. P. 152–160.
- Jämsä J., Ala-Kokko T., Huotari V. et al. Neutrophil CD64, C-reactive protein, and procalcitonin in the identification of sepsis in the ICU Post-test probabilities. // Journal of critical care. 2017. V. 43. P. 139–142.
- Kolaczkowska E., Kubes P. Neutrophil recruitment and function in health and inflammation // Nature Reviews Immunology. 2013. V. 13. № 3. P. 159–175.
- McEver R.P. Selectins: initiators of leucocyte adhesion and signalling at the vascular wall // Cardiovascular research. 2015. V. 107. № 3. P. 331–339.
- Miralda I., Uriarte S.M., McLeish K.R. Multiple Phenotypic Changes Define Neutrophil Priming // Frontiers in Cellular and Infection Microbiology. 2017. V. 7. P. 217.
- Ran T., Geng S., Li L. Neutrophil programming dynamics and its disease relevance // Science China Life Sciences. 2017.
- Rosales C. Fcγ Receptor Heterogeneity in Leukocyte Functional Responses // Frontiers in Immunology. 2017. V. 8.P. 280.
- Sundqvist M., Wekell P., Osla V. et al. Increased Intracellular Oxygen Radical Production in Neutrophils During Febrile Episodes of Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Cervical Adenitis Syndrome // Arthritis & Rheumatism. 2013. V. 65. № 11. P. 2971–2983.
- Tan S.-Y., Weninger W. Neutrophil migration in inflammation: intercellular signal relay and crosstalk // Current Opinion in Immunology. 2017. V. 44. P. 34–42.
- Zhang Y., Li W., Zhou Y. et al. Detection of sepsis in patient blood samples using CD64 expression in a microfluidic cell separation device // The Analyst. 2017. V. 143. № 1. P. 241–249.