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Cationic Liposomes Cause ROS Generation and Release of Neutrophil Extracellular Traps

  • N. Y. LotoshEmail author
  • S. O. Aliaseva
  • I. K. Malashenkova
  • G. M. Sorokoumova
  • R. G. Vasilov
  • A. A. SelischevaEmail author
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Abstract

Cationic liposomes are used as nanocarriers of drugs in chemotherapy, antibacterial therapy, and gene therapy. Positively charged stearylamine is commonly used in cationic liposome production. It has been shown that cationic liposomes, when applied intravenously, impact the functions and viability of blood cells, including neutrophils (innate immunity cells). In this work we studied the influence of phosphatidylcholine cationic liposomes containing stearylamine (SA liposomes) on the activity of human neutrophils–their ability of the formation of extracellular traps (NETs) and of the production of reactive oxygen species (ROS). The ratio of phosphatidylcholine to stearylamine in the liposomes was 9 : 1. Phosphatidylcholine liposomes (PC liposomes) were used as control. Liposomes were 140 ± 49 nm in diameter and differed in zeta-potential: PC liposomes had –1.74 ± 0.31 mV, while SA-liposomes had 11.40 ± 0.44 mV. NETs were visualized by light and fluorescent microscopy. It was found that the cells remained intact after a 90-min incubation with PC liposomes (control experiment), while incubation with SA liposomes caused an extrusion of thin DNA fibers into the extracellular space, i.e., the formation of NETs. ROS generation by neutrophils incubated with the liposomes and stimulated either with 0.5 mg/mL of zymozan, or with 40 nM of phorbol-12-miristate-13 acetate, or without stimulation was assessed by luminol-dependent chemiluminescence. It was shown that PC liposomes did not exert any significant effect on chemiluminescence of unstimulated and zymozan-stimulated neutrophils. PMA in the presence of liposomes did not exert any effect on the cells. SA liposomes had a complex impact on the activity of neutrophils: they inhibited the effects of zymozan and caused a prominent respiratory burst independently of the presence of the additional stimulation. The time before the maximal intensity of chemiluminescence peak after incubation with SA-liposomes ranged from 1 to 3 h and significantly differed from the time before the maximal intensity of chemoluminescence peak after incubation with zymozan or PMA (30–40 min). Apocynin, a NADPH-oxidase inhibitor, suppressed the respiratory burst induced by SA liposomes, indicating that NADPH oxidase has a role in ROS production caused by SA liposomes. To assess characteristics of the respiratory burst, lucigenin-dependent chemiluminescence sensitive to superoxide anion radical (\({\text{O}}_{2}^{{ \bullet -}}\)) was registered, and it was found that \({\text{O}}_{2}^{{ \bullet -}}\) is not produced after stimulation of neutrophils by SA liposomes. On the basis of these findings we conclude that SA liposomes are a polyfunctional factor that can affect the formation of NETs, suppress the action of zymozan, and stimulate ROS production by neutrophils.

Keywords:

chemiluminescence liposomes luminol NETs neutrophils phorbol ether respiratory burst stearylamine zymozan 

Notes

ACKNOWLEDGMENTS

Authors thank colleagues from MSU Biology department G.E. Onishchenko and N.V. Vorobjeva for valuable advice and consultations. The work was performed using the equipment of the Resource Center of the optical microscopy and spectroscopy; Resource Center of molecular and cell biology, and Resource Center of cognitive research of the Scientific Investigation Center “Kurchatovskii Institute”.

COMPLIANCE WITH ETHICAL STANDARDS

Conflict of interest. The authors declare that they have no conflicts of interest.

Ethical statement. 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. Informed consent was obtained from all individual participants involved in the study. This article does not contain any studies involving animals performed by any of the authors.

REFERENCES

  1. 1.
    Shvets V.I., Krasnopolskii Yu.M., Sorokoumova G.M. 2017. Liposomalnye formy lekarstvennykh preparatov (Liposomal forms of medicinal drugs). Moscow: Remedium.Google Scholar
  2. 2.
    Degim Z., Degim T., Bas L., Elmas M. 2002. The use of liposomal enrofloxacin for intracellular infections in Kangal dogs and visualization of phagocytosis of liposomes. J. Biomed. Mater. Res. 61, 246–251.CrossRefGoogle Scholar
  3. 3.
    Mitchell M.J., Wayne E., Rana K., Schaffer C.B., King M.R. 2014. TRAIL-coated leukocytes that kill cancer cells in the circulation. Proc. Natl Acad. Sci. USA. 111 (3), 930–935.CrossRefGoogle Scholar
  4. 4.
    Francian A., MannK., Kullberg M. 2017. Complement C3-dependent uptake of targeted liposomes into human macrophages, B cells, dendritic cells, neutrophils, and MDSCs. Intern. J. Nanomedicine. 12, 5149–5161.CrossRefGoogle Scholar
  5. 5.
    Sharma S., Rajendran V., Kulshreshtha R., Ghosh P.C. 2017. Enhanced efficacy of anti-miR-191 delivery throughstearylamine liposome formulation for the treatment of breast cancer cells. Int. J. Pharm. 15, 530 (1–2), 387–400. doi  https://doi.org/10.1016/j.ijpharm.2017.07.079
  6. 6.
    Sun J., Song Y., Lu M., Lin X., Liu Y., Zhou S., Su Y., Deng Y. 2016. Evaluation of the antitumor effect of dexamethasone palmitate and doxorubicin co-loaded liposomes modified with a sialic acid-octadecylamine conjugate. Eur. J. Pharm. Sci. 93, 177–183.CrossRefGoogle Scholar
  7. 7.
    Roychoudhury J., Sinha R., Ali N. 2011 Therapy with sodium stibogluconate in stearylamine-bearing liposomes confers cure against SSG-resistant Leishmania donovani in BALB/c mice. PLoS One. 6 (3), 7376. doi  https://doi.org/10.1371/journal.pone.0017376 CrossRefGoogle Scholar
  8. 8.
    Banerjee A., De M., Ali N. 2011. Combination therapy with paromomycin-associated stearylamine-bearing liposomes cures experimental visceral leishmaniasis through Th1-biased immunomodulation. Antimicrob. Agents Chemother. 55 (4), 1661–1670.CrossRefGoogle Scholar
  9. 9.
    Rajendran V., Rohra S., Raza M., Hasan G.M., Dutt S., Ghosh P.C. 2015. Stearylamine liposomal delivery of monensin in combination with free artemisinin eliminates blood stages of Plasmodium falciparum in culture and P. berghei infection in murine malaria. Antimicrob. Agents Chemother. 60 (3), 1304–1318. doi  https://doi.org/10.1128/AAC.01796-15 CrossRefGoogle Scholar
  10. 10.
    Hasan G.M., Garg N., Dogra E., Surolia R. Ghosh P.C. 2011. Inhibition of the growth of Plasmodium falciparum in culture by stearylamine-phosphatidylcholine liposomes. J. Parasitol. Res. 120 462. doi  https://doi.org/10.1155/2011/120462
  11. 11.
    Hwang T.L., Hsu C.Y., Aljuffali I.A., Chen C.H., Chang Y.T., Fang J.Y. 2015. Cationic liposomes evoke proinflammatory mediator release and neutrophil extracellular traps (NETs) toward human neutrophils. Colloids. Surf. B. Biointerfaces. 128, 119–126.CrossRefGoogle Scholar
  12. 12.
    Takano K., Sato K., Negishi Y., Aramaki Y. 2012. Involvement of actin cytoskeleton in macrophage apoptosis induced by cationic liposomes. Arch. Biochem. Biophys. 518 (1), 89–94.CrossRefGoogle Scholar
  13. 13.
    Arisaka M., Takano K., Negishi Y., Arima H., Aramaki Y. 2011. Involvement of lipid rafts in macrophage apoptosis induced by cationic liposomes. Arch. Biochem. Biophys. 508, 72–77.CrossRefGoogle Scholar
  14. 14.
    Arisaka M., Nakamura T., Yamada A., Negishi Y., Aramaki Y. 2010. Involvement of protein kinase C delta in induction of apoptosis by cationic liposomes in macrophage-like RAW264.7 cells. FEBS Lett. 584 (5), 1016–1020.CrossRefGoogle Scholar
  15. 15.
    Brinkmann, V., Reichard, U., Goosmann C., Fauler B., Uhlemann Y., Weiss D.S., Weinrauch Y., Zychlinsky A. 2004. Neutrophil extracellular traps kill bacteria. Science. 303, 1532–1535.CrossRefGoogle Scholar
  16. 16.
    Hwang T.L., Aljuffali I.A., Hung C.F., Chen C.H., Fang J.Y. 2015. The impact of cationic solid lipid nanoparticles on human neutrophil activation and formation of neutrophil extracellular traps (NETs). Chem. Biol. Interact. 235, 106–114.CrossRefGoogle Scholar
  17. 17.
    Sawai T., Asada M., Nishizawa Y., Nunoi H., Katayama K. 1999. Inhibition by alkylamines of NADPH oxidase through blocking the assembly of enzyme components. Jpn. J. Pharmacol. 80 (3), 237–242.CrossRefGoogle Scholar
  18. 18.
    Lotosh N.Yu., Moskalenko A.D., Malashenkova I.K., Kazanova G.V., Shchelkonogov V.A., Sorokoumova G.M., Vantsyan M.A., Selishcheva A.A., Vasilov R.G. 2017. Effects of liposomes of different composition on the oxidative burst in human neutrophils. Vestnik Biotekhnol. Fiz.-khim. Biol. im. Yu.A. Ovchinnikova (Rus.). 13 (1), 13–17.Google Scholar
  19. 19.
    Malech H.L., DeLeo F.R., Quinn M.T. 2012. The role of neutrophils in the immune system: An overview. In: Neutrophil Methods and Protocols. Ed. Quinn M.T. New York City: Humana Press, p. 3–13.Google Scholar
  20. 20.
    Makni-Maalej K., Chiandotto M., Hurtado-Nedelec M., Bedouhene S., Gougerot-Pocidalo M.A., Dang P.M., El-Benna J. 2013. Zymosan induces NADPH oxidase activation in human neutrophils by inducing the phosphorylation of p47phox and the activation of Rac2: Involvement of protein tyrosine kinases, PI3Kinase, PKC, ERK1/2 and p38MAPkinase. Biochem. Pharmacol. 85 (1), 92–100. doi  https://doi.org/10.1016/j.bcp.2012.10.010 CrossRefGoogle Scholar
  21. 21.
    Sumimoto H. 2008. Structure, regulation and evolution of Nox-family NADPH oxidases that produce reactive oxygen species. FEBS J. 275 (13), 3249–3277. doi  https://doi.org/10.1111/j.1742-4658.2008.06488.x CrossRefGoogle Scholar
  22. 22.
    Bedard K., Krause K.H. 2007. The NOX family of ROS-generating NADPH oxidases: Physiology and pathophysiology. Physiol. Rev. 87 (1), 245–313.CrossRefGoogle Scholar
  23. 23.
    Vorobjeva N.V., Pinegin B.V. 2016. Effects of the antioxidants Trolox, Tiron and Tempol on neutrophil extracellular trap formation. Immunobiology. 221 (2), 208–219.CrossRefGoogle Scholar
  24. 24.
    Vorobjeva N., Prikhodko A., Galkin I., Pletjushkina O., Zinovkin R., Sud’ina G., Chernyak B., Pinegin B. 2017. Mitochondrial reactive oxygen species are involved in chemoattractant-induced oxidative burst and degranulation of human neutrophils in vitro. Eur. J. Cell. Biol. 96 (3), 254–265. doi  https://doi.org/10.1016/j.ejcb.2017.03.003 CrossRefGoogle Scholar
  25. 25.
    Vladimirov Yu.A. 2001. Activated chemiluminescence and bioluminescence as a tool in medical and biological investigations. Sorosovskii obrazovatelnyi zhurnal (Rus.). 7 (1), 16–23.Google Scholar
  26. 26.
    Shchelkonogov V.A., Sorokoumova G.M., Baranova O.A., Chekanov A.V., Klochkova A.V., Kazarinov K.D., Solovieva E.Y., Fedin A.I., Shvets V.I. 2016. Liposomal form of lipoic acid: Preparation and determination of antiplatelet and antioxidant activity. Biomed. Khim. 62 (5), 577–583.CrossRefGoogle Scholar
  27. 27.
    Gavella M., Kveder M., Lipovac V. 2010. Modulation of ROS production in human leukocytes by ganglioside micelles. Braz. J. Med. Biol. Res. 43 (10), 942–949.CrossRefGoogle Scholar
  28. 28.
    Hosseini H., Li Y., Kanellakis P., Tay C., Cao A., Tipping P., Bobik A., Toh B.H., Kyaw T. 2015. Phosphatidylserine liposomes mimic apoptotic cells to attenuate atherosclerosis by expanding polyreactive IgM producing B1a lymphocytes. Cardiovasc. Res. 106 (3), 443–452.CrossRefGoogle Scholar
  29. 29.
    Remijsen Q. 2011. Neutrophil extracellular trap cell death requires both autophagy and superoxide generation. Cell Res. 21, 290–304CrossRefGoogle Scholar
  30. 30.
    de Buhr N., von Köckritz-Blickwede M. 2016. How neutrophil extracellular traps become visible. J. Immunol. Res. 2016, 460–473.CrossRefGoogle Scholar
  31. 31.
    Dikalov S. 2011. Crosstalk between mitochondria and NADPH oxidases. Free Radic. Biol. Med. 51 (7), 1289–1301.CrossRefGoogle Scholar
  32. 32.
    Kröller-Schön S., Steven S., Kossmann S., Scholz A., Daub S., Oelze M., Xia N., Hausding M., Mikhed Y., Zinssius E., Mader M., Stamm P., Treiber N., Scharffetter-Kochanek K., Li H., Schulz E., Wenzel P., Münzel T., Daibe A. 2014. Molecular mechanisms of the crosstalk between mitochondria and NADPH oxidase through reactive oxygen species-studies in white blood cells and in animal models. Antioxid. Redox Signal. 20 (2), 247–266.CrossRefGoogle Scholar
  33. 33.
    Dahlgren C., Karlsson A. 2002. Ionomycin-induced neutrophil NADPH oxidase activity is selectively inhibited by the serine protease inhibitor diisopropylfluorophosphate. Antioxid. Redox. Signal. 4 (1), 17–25.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2019

Authors and Affiliations

  • N. Y. Lotosh
    • 1
    • 2
    Email author
  • S. O. Aliaseva
    • 3
  • I. K. Malashenkova
    • 1
    • 4
  • G. M. Sorokoumova
    • 3
  • R. G. Vasilov
    • 1
  • A. A. Selischeva
    • 1
    • 2
    Email author
  1. 1.National Research Center “Kurchatov Institute”MoscowRussia
  2. 2.Moscow Lomonosov State University, Faculty of BiologyMoscowRussia
  3. 3.Moscow Technological University, Lomonosov Institute of Thin Chemical TechnologyMoscowRussia
  4. 4.Scientific Clinical Center of Physico-chemical MedicineMoscowRussia

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