Peculiarities of Phagocytosis of Opsonized and Nonopsonized Bacteria S. Aureus and E. Coli by Human Neutrophil Granulocytes Studied by Atomic Force Microscopy
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Differences in the phagocytosis process of opsonized and nonopsonized strains of Staphylococcus aureus 2879 M and Escherichia coli 321 were studied. Differences in the character of pseudopodia during phagocytosis by neutrophil granulocytes (NGs) of opsonized and nonopsonized bacteria were detected, and differences in the nature of pseudopodia in reactions to gram-positive and gram-negative microorganisms were not detected. For the first time in dynamic observations at the late stages of phagocytosis, changes in the volume of nuclei and their movement, variations in the intersegment distance of the nuclei, and a slight increase in the volume of NGs were shown. A decrease in the rigidity of the membrane–cytoskeleton NG complex correlating with the intensity of phagocytosis and opsonization of bacteria was shown for the first time. It was established that opsonization does not affect the oxygen-dependent metabolism of NGs and, at the same time, introduces significant adjustments in the implementation of oxygen-independent bactericidal mechanisms of cells.
Keywords:neutrophil granulocytes phagocytosis opsonization S. aureus E. coli pseudopodia atomic force microscopy rigidity of the membrane–cytoskeleton complex
This study was supported by the Russian Science Foundation, project no. 16-14-10179.
COMPLIANCE WITH ETHICAL STANDARDS
Сonflict of interests. The authors declare that they have no conflict of interest.
Statement on the welfare of animals. This article does not contain any studies with animals performed by any of the authors.
Statement of compliance with standards of research involving humans as subjects. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Blood sampling was performed in the morning, after the donors signed the consent form.
- 1.Ackerman, G.A., A modification of the Sudan Black B technique for the possible cytochemical demonstration of masked lipids, J. Natl. Cancer Inst., 1952, vol. 13, pp. 219–220.Google Scholar
- 3.Belotskii, S.M. and Avtalion, R.R., Vospalenie, mobilizatsiya kletok i klinicheskie effekty (Inflammation, Cell Mobilization and Clinical Effects), Moscow: Binom, 2008.Google Scholar
- 5.Bukharaev, A.A., Mozhanova, A.A., Nurgazizov, N.I., and Ovchinnikov, D.V., Measuring local elastic properties of cell surfaces and soft materials in liquid by atomic force microscopy, Phys. Low-Dimens. Struct., 2003, vols. 3–4, pp. 31–38.Google Scholar
- 6.Burstone, M.S., Histochemical demonstration of acid phosphatases with naphthol AS-phosphates, J. Natl. Cancer Inst., 1958, vol. 21, pp. 523–539.Google Scholar
- 8.Fritz-Laylin, L.K., Riel-Mehan, M., Chen, B.C., Lord, S.J., Goddard, T.D., Ferrin, T.E., Nicholson-Dykstra, S.M., Higgs, H., Johnson, G.T., Betzig, E., and Mullins, R.D., Actin-based protrusions of migrating neutrophils are intrinsically lamellar and facilitate direction changes, Elife, 2017. doi 10.7554/eLife.26990Google Scholar
- 9.Hayhoe, F.G.J. and Quaglino, D., Haematological Cytochemistry, Edinburgh: Churchill Livingstone, 1980. 336 p.Google Scholar
- 10.Leithner, A., Eichner, A., Müller, J., Reversat, A., Brown, M., Schwarz, J., Merrin, J., de Gorter, D.J., Schur, F., Bayerl, J., de Vries, I., Wieser, S., Hauschild, R., Lai, F.P., Moser, M., Kerjaschki, D., Rottner, K., Small, J.V., Stradal, T.E., and Sixt, M., Diversified actin protrusions promote environmental exploration but are dispensable for locomotion of leukocytes, Nat. Cell Biol., 2016, vol. 18, pp. 1253–1259.CrossRefGoogle Scholar
- 11.Loomis, W.F., Fuller, D., Gutierrez, E., Groisman, A., and Rappel, W.J., Innate non-specific cell substratum adhesion, PLoS One, 2012, vol. 7. doi 10.1371/journal.pone.0042033Google Scholar
- 13.Mayanskii, A.N., Patogeneticheskaya mikrobiologiya (Pathogenic Microbiology), Nizhny Novgorod: Izd. Nizhegorod. Gos. Med. Akad., 2006.Google Scholar
- 14.Mayanskii, A.N. and Pikusa, O.I., Klinicheskie aspekty fagotsitoza (The Clinical Aspects of Phagocytosis), Kazan: Magarif, 1993.Google Scholar
- 17.Pigarevskii, V.E., Klinicheskaya morfologiya neitrofil’nykh granulotsitov (The Clinical Morphology of Neutrophil Granulocytes), Leningrad: Nauka, 1988.Google Scholar
- 18.Pleskova, S.N., Atomno-silovaya mikroskopiya v biologicheskikh i meditsinskikh issledovaniyakh (Atomic-Force Microscopy in Biology and Medicine), Dolgoprudnyi: Intellekt, 2011.Google Scholar
- 19.Pleskova, S.N., Guschina, Yu.Yu., and Zvonkova, M.B., Investigation of the influence of complement system on the various strains of proteus by methods of atomic force microscopy and luminol-dependent chemiluminescence, Phys. Low-Dimens. Struct, 2004, vols. 1–2, pp. 77–82.Google Scholar
- 20.Pleskova, S.N., Zvonkova, M.B., and Gushchina, Yu.Yu., Studying the neutrophil granulocytes morphological characteristics by scanning probe microscopy. Morphology, Arkh. Anat. Gistol. Embriol., 2005, vol. 127, no. 1, pp. 60–62.Google Scholar
- 21.Podosinnikov, I.S., Nilova, L.G., Babichenko, I.V., Turina, O.P., and Ponomareva, V.N., Method for determining the chemotactic activity of leukocytes, Lab. Delo, 1981, vol. 8, pp. 468–470.Google Scholar
- 24.Shubich, M.G. and Nagoev, B.S., Shchelochnaya fosfataza leikotsitov v norme i patologii (Alkaline Phosphatase of Leukocytes in Norm and Pathology), Moscow: Meditsina, 1980.Google Scholar
- 25.Van Kessel, K.P., Bestebroer, J., and van Strijp, J.A., Neutrophil-mediated phagocytosis of Staphylococcus aureus, Front. Immunol., 2014, vol. 5. doi 10.3389/fimmu.2014.00467Google Scholar
- 26.Vandenbroucke-Grauls, C.M., Thijssen, H.M., and Verhoef, J., Opsonization of Staphylococcus aureus protects endothelial cells from damage by phagocytosing polymorphonuclear leukocytes, Infect. Immun., 1987, vol. 55, pp. 1455–1460.Google Scholar
- 27.Vargas, P., Maiuri, P., Bretou, M., Sáez, P.J., Pierobon, P., Maurin, M., Chabaud, M., Lankar, D., Obino, D., Terriac, E., Raab, M., Thiam, H.R., Brocker, T., Kitchen-Goosen, S.M., Alberts, A.S., Sunareni, P., Xia, S., Li, R., Voituriez, R., Piel, M., and Lennon-Duménil, A.M., Innate control of actin nucleation determines two distinct migration behaviours in dendritic cells, Nat. Cell Biol., 2016, vol. 18, pp. 43–53.CrossRefGoogle Scholar