Dose and Time-Dependent Lipopolysaccharide Exposure on A549 Cell Model Influences Pro-Inflammatory Cytokine Interleukin 8

Sarah Sumayyah, Prayoga Suryadarma, Rachmawati Noverina, Wireni Ayuningtyas, Firman Fuad Wirakusumah, Ahmad Faried

Abstract


Hyperinflammation in COVID-19 patients is one of the causes of the high mortality rate of COVID-19. An in vitro model mimicking the inflammatory responses in COVID-19 patients is important in the efforts of finding new drug candidates for this disease. Lipopolysaccharide (LPS) can increase the proinflammatory cytokine interleukin 8 in response to the presence of foreign substances. This preliminary study sought to explore the use of  the A549 cells as an in vitro inflammatory model. This study was conducted from August to November 2022 at the stem cell research and development laboratory of Bio Farma Indonesia. The exposure of 100, 500, and 1000 g/mL doses of LPS administered for 24, 72, and 120 hours on the A549 cells was analyzed for cell viability, population doubling time (PDT), and the presence of  proinflammatory cytokine IL-8. The group differences were examined using one- and two-way analysis of variance in IBM SPSS Statistics Version 29, with a p-value of 0.05 considered significant. Cells exposed to a dose of 1000 g/mL LPS had a lower viability and a higher proliferation rate (p<0.05) based on the viability and PDT. Viability, PDT, and pro-inflammatory cytokines showed concentration- and time-dependent responses. Therefore, increased levels of the proinflammatory cytokine IL-8 in cells exposed to LPS at a dose of 1000 g/mL for 24 hours can be used as a mimic to study hyperinflammation in COVID-19 patients.

Keywords


A549 cell, inflammation, interleukin 8, lipopolysaccharide exposure

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  1. Darif D, Hammi I, Kihel A, el Idrissi Saik I, Guessous F, Akarid K. The pro-inflammatory cytokines in COVID-19 pathogenesis: What goes wrong?. Microb Pathog. 2021;153:104799.
  2. Henderson LA, Canna SW, Schulert GS, et al. On the Alert for Cytokine Storm: Immunopathology in COVID-19. Arthritis and Rheumatology. 2020;72(7):1059–63.
  3. Al-Ani B, ShamsEldeen AM, Kamar SS, Haidara MA, Al-Hashem F, Alshahrani MY. Lipopolysaccharide induces acute lung injury and alveolar haemorrhage in association with the cytokine storm, coagulopathy and AT1R/JAK/STAT augmentation in a rat model that mimics moderate and severe Covid-19 pathology. Clin Exp Pharmacol Physiol. 2022;49(4):483–91.
  4. von Schéele I, Larsson K, Palmberg L. Interactions between alveolar epithelial cells and neutrophils under pro-inflammatory conditions. Eur Clin Respir J. 2014;1(1):24545.
  5. Kuwano K, Yanagihara T, Hamada N, et al. Amphiregulin suppresses epithelial cell apoptosis in lipopolysaccharide-induced lung injury in mice. Biochem Biophys Res Commun. 2017;484(2):422–8.
  6. Nova Z, Skovierova H, Strnadel J, Halasova E, Calkovska A. Short-term versus long-term culture of A549 cells for evaluating the effects of lipopolysaccharide on oxidative stress, surfactant proteins and cathelicidin LL-37. Int J Mol Sci. 2020;21(3):1148.
  7. Thorley AJ, Ford PA, Giembycz MA, Goldstraw P, Young A, Tetley TD. Differential regulation of cytokine release and leukocyte migration by lipopolysaccharide-stimulated primary human lung alveolar type II epithelial cells and macrophages. J Immunol. 2007;178(1):463–73.
  8. Fuchs S, Hollins AJ, Laue M, Schaefer UF, Roemer K, Gumbleton M, et al. Differentiation of human alveolar epithelial cells in primary culture: Morphological characterization and synthesis of caveolin-1 and surfactant protein-C. Cell Tissue Res. 2003;311(1):31–45.
  9. Cooper JR, Abdullatif MB, Burnett EC, Kempsell KE, Conforti F, Tolley H, et al. Long term culture of the a549 cancer cell line promotes multilamellar body formation and differentiation towards an alveolar type II Pneumocyte phenotype. PLoS One. 2016;11(10):e0164438.
  10. Nova Z, Skovierova H, Calkovska A. Alveolar-capillary membrane-related pulmonary cells as a target in endotoxin-induced acute lung injury. Int J Mol Sci. 2019;20(4):831.
  11. Li C, Ma D, Zhang M, An L, Wu C, Zhou H. Effects of long-time exposure to lipopolysaccharide on intestinal lymph node immune cells and antibodies level in mice. Iranian J Immunol. 2020;17(3):175–84.
  12. Cesta MC, Zippoli M, Marsiglia C, et al. The role of interleukin-8 in lung inflammation and injury: implications for the management of COVID-19 and hyperinflammatory acute respiratory distress syndrome. Front Pharmacol. 2022;12:808797.
  13. Wiesmann N, Gieringer R, Viel M, Eckrich J, Tremel W, Brieger J. Zinc oxide nanoparticles can intervene in radiation-induced senescence and eradicate residual tumor cells. Cancers (Basel). 2021;13(12):2989.
  14. McElvaney OJ, McEvoy NL, McElvaney OF, Carroll TP, Murphy MP, Dunlea DM, et al. Characterization of the inflammatory response to severe COVID-19 Illness. Am J Respir Crit Care Med. 2020;202(6):812–21.
  15. Long X, Ye Y, Zhang L, Liu P, Yu W, Wei F, et al. IL-8, a novel messenger to cross-link inflammation and tumor EMT via autocrine and paracrine pathways (Review). Int J Oncol. 2016;48(1):5–12.
  16. Li L, Li J, Gao M, Fan H, Wang Y, Xu X, et al. Interleukin-8 as a biomarker for disease prognosis of coronavirus disease-2019 Patients. Front Immunol. 2021;11:602395.
  17. Zhao J, Li X, Zou M, He J, Han Y, Wu D, et al. MiR-135a inhibition protects A549 cells from LPS-induced apoptosis by targeting Bcl-2. Biochem Biophys Res Commun. 2014;452(4):951–7.
  18. Li S, Guo L, Qian P, Zhao Y, Liu A, Ji F, Chen L, et al. Lipopolysaccharide induces autophagic cell death through the PERK-dependent branch of the unfolded protein response in human alveolar epithelial A549 cells. Cell Physiol Biochem. 2015;36(6):2403–17.
  19. Migale R, Herbert BR, Lee YS, Sykes L, Waddington SN, Peebles D, et al. Specific lipopolysaccharide serotypes induce differential maternal and neonatal inflammatory responses in a murine model of preterm labor. Am J Pathol. 2015;185(9):2390–401.
  20. Eder K, Vizler C, Kusz E, Karcagi I, Glavinas H, Balogh GE, et al. The role of lipopolysaccharide moieties in macrophage response to Escherichia coli. Biochem Biophys Res Commun. 2009;389(1):46–51.
  21. Chen L, Wang G, Tan J, Cao Y, Long X, Luo H, et al. Scoring cytokine storm by the levels of MCP-3 and IL-8 accurately distinguished COVID-19 patients with high mortality. Signal Transduct Target Ther. 2020;5(1):292.
  22. Bendib I, Beldi-Ferchiou A, Schlemmer F, Surenaud M, Maitre B, Plonquet A, et al. Alveolar compartmentalization of inflammatory and immune cell biomarkers in pneumonia-related ARDS. Crit Care. 2021;25(1):23.
  23. Chiang CC, Korinek M, Cheng WJ, Hwang TL. Targeting neutrophils to treat acute respiratory distress syndrome in coronavirus disease. Front Pharmacol. 2020;11:572009




DOI: https://doi.org/10.15395/mkb.v55n2.3143

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