Makrofajlarda Neopterin ve IP-10 Üretimine İnterlökin-33’ün Etkileri

Year 2023, Volume: 76 Issue: 1, 30 - 36, 25.05.2023

Rahime Aksoy Bulgurcu , Cemalettin Aybay , Hüseyin Tutkak , Vedat Bulut

Abstract

Amaç: İnterleukin-33 (IL-33), IL-1 ailesinden pleiotropik bir sitokindir. Th2 yönünde plastisiteye yol açtığı bilinmektedir. Doğal bağışıklık sisteminin önemli hücreleri olan makrofajlar, IL-33 reseptörü ST2L’yi ifade eder ve IL-33 etkisi altında M2 makrofaj polaritesi sergiler. Makrofajlar, interferon-γ ile indüklenen bir kemokin olan interferon gama ile uyarılan protein 10 (IP-10) ve hücresel immünoreaksiyon gösteren neopterin üretir. Bu çalışmanın amacı, uyarılmamış ve farklı sitokinlerle önceden uyarılmış makrofajlarda IL-33’ün neopterin ve IP-10 üretimi üzerindeki etkilerini araştırmaktır. Gereç ve Yöntem: J774.1 makrofaj hücre dizisi kullanıldı. J774.1 makrofajları, IL-33 (100 ng/mL) ile ön işlemden sonra IFN-γ (40 U/mL) + LPS (10 ng/mL) ile uyarıldı. Nitrit seviyeleri Griess reaksiyonu ile ölçüldü. IL-33’ün polarize makrofajlar üzerindeki etkisini incelemek için J774.1 makrofajları, M1, M2a ve M2c makrofaj polarizasyonu sağlamak üzere IL-4, IL-10 ve IL-13 sitokinleri ile uyarıldı. Neopterin ve IP-10 ölçümleri ELISA yöntemi ile yapıldı. İstatistiksel analiz SPSS 18 yazılımı ile yapıldı. Bulgular: IL-33’ün, uyarılmamış ve plastisitesi etkilenmiş J774.1 hücrelerinde neopterin üretimini önemli ölçüde artırdığı bulundu. Tüm deneylerde IP-10 seviyelerinde değişiklik olmadı. Neopterin ve IP-10 seviyeleri arasında bir korelasyon bulunmadı. Sonuç: Bu çalışmamız, IL-33 sitokinin neopterin üretimi üzerindeki etkileri üzerine yapılan ilk çalışmadır. IL-33, J774.1 hücrelerinde neopterin üretiminde önemli artışa neden olmaktadır.

Keywords

İnterlökin-33 , IP-10 , Neopterin , Makrofaj , ELİZA

Ethical Statement

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Effects of Interleukin-33 on Neopterin and IP-10 Production in Macrophages

Abstract

Objectives: Interleukin-33 (IL-33) is a pleiotropic cytokine from the IL-1 family. It is known to cause plasticity in the Th2 direction. The macrophages, important cells of the innate immune system, express the IL-33 receptor ST2L and exhibit M2 macrophage polarity under the influence of IL-33. Macrophages produce interferon gamma-stimulated protein 10 (IP-10), an interferon-γ-induced chemokine, and neopterin which exhibits cellular immunoreaction. The aim of this study was to investigate the effects of IL-33 on neopterin and IP-10 production in unstimulated and prestimulated macrophages with different cytokines. Materials and Methods: The J774.1 macrophage cell line was used. J774.1 macrophages were stimulated by IFN-γ (40 U/mL) + LPS (10 ng/mL) after pretreatment with IL-33 (100 ng/mL). Nitrit levels were measured by the Griess reaction. To examine the effect of IL-33 on polarized macrophages, J774.1 macrophages were stimulated with IL-4, IL-10 ve IL-13 cytokines to provide M1, M2a and M2c macrophage polarization. Neopterin and IP-10 measurements were carried out by ELISA method. Statistical analysis was performed with SPSS 18 software. Results: IL-33 was found to significantly increase neopterin production in unstimulated and plasticity-affected J774.1 cells. There was no change in IP-10 levels in all experiments. No correlation was found between neopterin and IP-10 levels. Conclusion: Our present study is the first study on the effects of IL-33 cytokine on neopterin production. IL-33 causes a significant increase in neopterin production in J774.1 cells.

Keywords

Interleukin-33 , IP-10 , Neopterin , Macrophage , ELISA

Ethical Statement

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References

• 1. Zhang C, Yang M, Ericsson AC. Function of macrophages in disease: current understanding on molecular mechanisms. Front Immunol. 2021;12:620510.
• 2. Finlay CM, Cunningham KT, Doyle B, et al. IL-33-stimulated murine mast cells polarize alternatively activated macrophages, which suppress t cells that mediate experimental autoimmune encephalomyelitis. J Immunol. 2020;205:1909-1919.
• 3. Guo H, Bossila EA, Ma X, et al. Dual immune regulatory roles of interleukin-33 in pathological conditions. Cells. 2022;11:3237.
• 4. Schmitz J, Owyang A, Oldham E, et al. IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines. Immunity. 2005;23:479-490.
• 5. Cayrol C. IL-33, an alarmin of the il-1 family involved in allergic and non allergic inflammation: focus on the mechanisms of regulation of its activity. Cells. 2021;11:107.
• 6. Lang S, Li L, Wang X, et al. J. CXCL10/IP-10 Neutralization can ameliorate lipopolysaccharide-induced acute respiratory distress syndrome in rats. PLoS One. 2017;12:e0169100.
• 7. Makuch E, Jasyk I, Kula A, et al. IFNβ-Induced CXCL10 chemokine expression is regulated by pellino3 ligase in monocytes and macrophages. Int J Mol Sci. 2022;23:14915.
• 8. Watanabe T. Neopterin derivatives - a novel therapeutic target rather than biomarker for atherosclerosis and related diseases. Vasa. 2021;50:165-173.
• 9. El-Hawy MA, Bedair HM, Madkour AMYM, et al. Assessment of serum neopterin and kynurenine levels in Egyptian children with sickle cell disease: a single center study. Pediatric Hematology/Oncology and Immunopathology. 2022;21:22-27.
• 10. Gieseg SP, Baxter-Parker G, Lindsay A. Neopterin, inflammation, and oxidative stress: what could we be missing? Antioxidants (Basel). 2018;7:80.
• 11. Peterbauer A, Eperon S, Jungi TW, et al. Interferon-gamma-primed monocytoid cell lines: optimizing their use for in vitro detection of bacterial pyrogens. J Immunol Methods. 2000;233:67-76.
• 12. Shanmuganathan MV, Krishnan S, Fu X, et al. Escherichia coli K1 induces pterin production for enhanced expression of Fcγ receptor I to invade RAW 264.7 macrophages. Microbes Infect. 2014;16:134-141.
• 13. Akhter N, Kochumon S, Hasan A, et al. IFN-γ and LPS Induce Synergistic Expression of CCL2 in Monocytic Cells via H3K27 Acetylation. J Inflamm Res. 2022 Jul 27;15:4291-4302.
• 14. Tighe RM, Liang J, Liu N, et al. Recruited exudative macrophages selectively produce CXCL10 after noninfectious lung injury. Am J Respir Cell Mol Biol. 2011;45:781-788.
• 15. Lyons C, Fernandes P, Fanning LJ, et al. Engagement of Fas on Macrophages Modulates Poly I:C induced cytokine production with specific enhancement of IP-10. PLoS One. 2015;10:e0123635.
• 16. Zhou Z, Yan F, Liu O. Interleukin (IL)-33: an orchestrator of immunity from host defence to tissue homeostasis. Clin Transl Immunology. 2020;9:e1146.
• 17. Tran VG, Cho HR, Kwon B. IL-33 priming enhances peritoneal macrophage activity in response to Candida albicans. Immune Netw. 2014;14:201-216.
• 18. Villarreal DO, Weiner DB. Interleukin 33: a switch-hitting cytokine. Curr Opin Immunol. 2014;28:102-106.
• 19. Jiang HR, Milovanović M, Allan D, et al. IL-33 attenuates EAE by suppressing IL-17 and IFN-γ production and inducing alternatively activated macrophages. Eur J Immunol. 2012;42:1804-1814.
• 20. Wu J, Li Q, Deng J, et al. Association between IL-33 and other inflammatory factors in patients with rheumatoid arthritis and in fibroblast-like synoviocytes in vitro. Exp Ther Med. 2021;21:161.
• 21. Tarique AA, Logan J, Thomas E, et al. Phenotypic, functional, and plasticity features of classical and alternatively activated human macrophages. Am J Respir Cell Mol Biol. 2015;53:676-688.
• 22. Kurowska-Stolarska M, Stolarski B, Kewin P, et al. IL-33 amplifies the polarization of alternatively activated macrophages that contribute to airway inflammation. J Immunol. 2009;183:6469-6477.
• 23. Mosser DM, Hamidzadeh K, Goncalves R. Macrophages and the maintenance of homeostasis. Cell Mol Immunol. 2021;18:579-587.