EXPLORING EFFECTS OF INTERLEUKIN-33 ON JAK/STAT SIGNALLING PATHWAYS IN MACROPHAGES

Year 2022, Volume: 4 Issue: 1, 32 - 44, 30.04.2022

Mehmet Zafer Kalayci , Zubeyir Elmazoglu , Vedat Bulut

Abstract

It is known that IL-33 cytokine plays role in pathogenesis of some diseases. The dual role of
macrophages within the tumor microenvironment is well-known and dependent on their plasticity.
Macrophages mainly serve as the component of innate immune system, but they also act as a bridge
between innate and acquired immune system with their antigen presentation capacity. Macrophages
can change their functions by being polarized to classical M1 and alternative M2 macrophages via
cytokine cocktails, such as interferon-, transforming growth factor- and interleukin-4, -10, -13. Tumor promoting effects are mainly associated with tumor-associated macrophages (TAMs)
resembling the M2 types of macrophages. On the other hand, macrophages shifted to M1 type are
capable of antitumour activities.
Investigation of down-stream intracellular signaling pathways is very important in order to
understand completely its role in pathogenesis of diseases mentioned above, as well as, to determine
appropriate hypothesis in studies of therapeutic agents related to down-stream intracellular signaling
pathways.
This study is aimed to enlighten the effect of IL-33 on tyrosine kinase and JAK/STAT intracellular
signaling pathways in macrophage cell line, J774.1. Therefore, mrIL-33 in several concentrations
will be added into cell cultures of J774 stimulated by IFN-γ plus LPS or unstimulated. Then, tyrosine
kinase (Tk) activities, levels and activation of Jak/STAT activities will be measured.
In this study, it was observed that IL-33 additions in prior to mrIFN-γ 40 U/mL plus LPS 10 ng/mL
stimulation decreased Tyk-2 levels in the group in which IL-33 levels were used at 10 ng/mL
(p<0.05). In order to monitor the effects of IL-33 on tyk2, it was observed that tyrosine kinase tyk2
levels did not change when IL-33 was added to cell cultures at 10ng/mL and 100 ng/mL
concentrations after 0, 4 and 18 hours. When IL-33 was applied to the J774.1 macrophage cell line at
a concentration of 10 ng/mL at different times, an increase in Tyk-2 phosphorylation was observed.
When IL-33 was applied to J774.1 cells at concentrations of 10 ng/mL and 100 ng/mL 4 and 18
hours before IFN-γ stimulation, statistically increased phosphorylated STAT levels were found in all
IL-33 applied groups compared to the control group. These results show that high-dose IL-33
administration creates a synergistic effect on STAT activation with IFN-γ stimulation.
This study will lead to observation of the effect of IL-33 in intracellular signaling pathways of
macrophages. Then, it will be going through the proper channels to understand effective mechanisms
of IL-33, as a therapeutic target in neoplasms.

Keywords

IL-33, intracellular signaling pathways, tyrosine kinase, JAK, STAT, INF-γ

References

• Referans1. Alves-Filho, J.C. et al., (2010). Interleukin-33 attenuates sepsis by enhancing neutrophil influx to the site of infection. Nature Medicine. 16 (6): p. 708-712.
• Referans2. Barrett, C.S., Millena A.C. and Khan S.A. (2017). TGF-β Effects on Prostate Cancer Cell Migration and Invasion Require FosB. Prostate 77 (1): p. 72-81.
• Referans3. Bilen, A. et al.,(2021). Does daily fasting shielding kidney on hyperglycemia-related inflammatory cytokine via TNF-α, NLRP3, TGF-β1 and VCAM-1 mRNA expression. International Journal of Biological Macromolecules. 190 : p. 911-918.
• Referans4. Biswas, S.K. et al.,(2012). Macrophage polarization and plasticity in health and disease. Immunologic Research. 53 (1): p. 11-24.
• Referans5. Boengler, K. et al.,(2008). The myocardial JAK/STAT pathway: from protection to failure. Pharmacology & Therapeutics. 120 (2): p. 172-185.
• Referans6. Bouff, C. et al.,(2013). IL-33 markedly activates murine eosinophils by an NF-κB–dependent mechanism differentially dependent upon an IL-4–driven autoinflammatory loop. The Journal of Immunology. 191 (8): p. 4317-4325.
• Referans7. Carlock, C.I. et al.,(2014). Unique temporal and spatial expression patterns of IL-33 in ovaries during ovulation and estrous cycle are associated with ovarian tissue homeostasis. The Journal of Immunology. 193 (1): p. 161-169.
• Referans8. Cassetta. L., Cassol E. and Poli G.,(2011). Macrophage polarization in health and disease. The Scientific World Journal. 11:p. 2391-2402.
• Referans9. Ciccia, F. et al.,(2013). IL-33 is overexpressed in the inflamed arteries of patients with giant cell arteritis. Annals of the rheumatic diseases. 72 (2): p. 258-264.
• referans10. Darnell, Jr J.E., (1997). STATs and gene regulation. Science. 277 (5332): p. 1630-1635.
• Referans11. Darnell, J.E.,Kerr I.M and Stark G.R, (1994).Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science. 264 (5164): p. 1415- 1421.
• Referans12. De Felice, F.G. and Ferreira S.T.,(2014).Inflammation, defective insulin signaling, and mitochondrial dysfunction as common molecular denominators connecting type 2 diabetes to Alzheimer's disease. Diabetes. 63 (7): p. 2262-2272. Referans13. DeNardo, D.G. and Ruffell, B. (2019). Macrophages as regulators of tumor immunity and immunotherapy. Nat Rev. Immunol. 19:369-382.
• Referans14.Drake, L.Y. and Kita H.,(2017). IL-33: biological properties, functions, and roles in airway disease. Immunological Reviews. 278 (1): p. 173-184.
• Referans15. Espinassous, Q. et al.,(2009). IL-33 enhances lipopolysaccharide-induced inflammatory cytokine production from mouse macrophages by regulating lipopolysaccharide receptor complex. The Journal of Immunology. 183 (2): p. 1446-1455.
• Referans16. Feghali, C.A. and Wright, T.M. (1997) Cytokines in acute and chronic inflammation. Front Biosci. 2 : p. d12-26.
• Referans17. Frankenberger. M., Pechumer H. and Ziegler-Heitbrock H., (1995). Interleukin-10 is upregulated in LPS tolerance. Journal of İnflammation. 45 (1): p. 56-63.
• Referans18. Fultz, M.J. et al., (1993). Induction of IFN-γ in macrophages by lipopolysaccharide. International Immunology. 5 (11): p. 1383-1392.
• Referans19. Goldring, M.B. and Goldring S.R.,(1991). Cytokines and cell growth control. Crit Rev. Eukaryote Gene Expr. 1 (4): p. 301-26.
• Referans20. Gordon, S. and Martinez F.O.,(2010). Alternative activation of macrophages: mechanism and functions. Immunity. 32 (5): p. 593-604.
• Referans21. Groner, B. and von Manstein V.,(2017). Jak Stat signaling and cancer: Opportunities, benefits and side effects of targeted inhibition. Molecular and cellular endocrinology. 451 : p. 1-14.
• Referans22. Heneka, M.T., Kummer M.P. and Latz E.,(2014). Innate immune activation in neurodegenerative disease. Nature Reviews Immunology. 14 (7): p. 463-477.
• Referans23. Hu X. et al.,(2007). Crosstalk among Jak ‐ STAT, Toll ‐ like receptor, and ITAM-dependent pathways in macrophage activation. Journal of leukocyte biology. 82 (2): p. 237-243.
• Referans24. Hudson C.A. et al.,(2008) Induction of IL-33 expression and activity in central nervous system glia. Journal of leukocyte biology. 84 (3): p. 631-643.
• Referans25. Jurkovich, G.J. et al., (1991). Interferon γ increases sensitivity to endotoxin. Journal of Surgical Research. 51 (3): p. 197-203.
• Referans26. Kakkar, R. and Lee R.T.,(2008). The IL-33/ST2 pathway: therapeutic target and novel biomarker. Nature reviews Drug discovery. 7 (10): p. 827-840.
• Referans27. Kearley, J. et al.,(2015). Cigarette smoke silences innate lymphoid cell function and facilitates an exacerbated type I interleukin-33-dependent response to infection. Immunity. 42 (3): p. 566-579.
• Referans28. Keskin, H. et al.,(2021) Neuroprotective effect of roflumilast under cerebral ischaemia/reperfusion injury in juvenile rats through NLRP-mediated inflammatory response inhibition. Clinical and Experimental Pharmacology and Physiology.48 (8): p. 1103-1110.
• Referans29. Kim S.K. et al.,(2011). Melittin enhances apoptosis through suppression of IL-6/sIL-6R complex- induced NF-κB and STAT3 activation and Bcl-2 expression for human fibroblast-like synoviocytes in rheumatoid arthritis. Joint Bone Spine.78 (5): p. 471-477.
• Referans30. Kurowska-Stolarska, M. et al.,(2009). IL-33 amplifies the polarization of alternatively activated macrophages that contribute to airway inflammation. The Journal of Immunology. 183 (10): p. 6469-6477.
• Referans31. Learn, C.A., Mizel, S.B. and McCall C.E,(2000). mRNA and protein stability regulates the differential expression of pro-and anti-inflammatory genes in endotoxin-tolerant THP-1 cells. Journal of Biological Chemistry. 275 (16): p. 12185-12193.
• Referans32. Li, D. et al.,(2014). IL-33 promotes ST2-dependent lung fibrosis by the induction of alternatively activated macrophages and innate lymphoid cells in mice. Journal of Allergy and Clinical Immunology. 134 (6): p. 1422-1432. e11.
• Referans33. Li, H.X. et al.,(2015). Retinoic acid amide inhibitors JAK/STAT pathway in lung cancer which leads to apoptosis. Tumor Biology. 36 (11): p. 8671-8678.
• Referans34. Liew, F.Y., Girard J.P. and Turnquist H.R.,(2016). Interleukin-33 in health and disease. Nature Reviews Immunology. 16 (11): p. 676-689.
• Referans35. Liongue, C., Sertori, R. and Ward A.C,(2016). Evolution of Cytokine Receptor Signaling. The Journal of Immunology. 197 (1): p. 11-18.
• Referans36. Lorsbach, R.B. et al.,(1993). Expression of the nitric oxide synthase gene in mouse macrophages activated for tumor cell killing. Molecular basis for the synergy between interferon-gamma and lipopolysaccharide. Journal of Biological Chemistry. 268 (3): p. 1908-1913.
• Referans37. Martin N.T. and Martin M.U.,(2016). Interleukin 33 is a guardian of barriers and a local alarm. Nature immunology. 17 (2): p. 122-131.
• Referans38. Mezouar, S. and Mege J.L.,(2020). Changing the paradigm of IFN-γ at the interface between innate and adaptive immunity: Macrophage ‐ derived IFN-γ. Journal of leukocyte biology. 108 (1): p. 419-426.
• Referans39. Mirchandani, A.S., Salmond, R.J. and Liew F.Y.,(2012). Interleukin-33 and the function of innate lymphoid cells. Trends in immunology. 33 (8): p. 389-396.
• Referans40. Molofsky, A.B., Savage A.K. and Locksley R.M.,(2015). Interleukin-33 in tissue homeostasis, injury, and inflammation. Immunity. 42 (6): p. 1005-1019.
• Referans41. Morris, R., Kershaw, N.J. and Babon J.J, (2018) The molecular details of cytokine signaling via the JAK/STAT pathway. Protein science:a publication of the Protein Society. 27 (12): p. 1984- 2009.
• Referans42. Moussion, C., Ortega, N. and Girard J.P.,(2008). The IL-1-like cytokine IL-33 is constitutively expressed in the nucleus of endothelial cells and epithelial cells in vivo: a novel 'alarmin'? PloS one. 3 (10): p. e3331.
• Referans43. Onda, H. et al.,(1999). Identification of genes differentially expressed in canine vasospastic cerebral arteries after subarachnoid hemorrhage. Journal of Cerebral Blood Flow & Metabolism. 19 (11): p. 1279-1288.
• Referans44. O'Shea, J.J. et al.,(2015). The JAK-STAT pathway: impact on human disease and therapeutic intervention. Annual review of medicine. 66 : p. 311-328.
• Referans45. O'Shea, J.J. and Plenge R.,(2012). JAK and STAT signaling molecules in immunoregulation and immune-mediated disease. Immunity. 36 (4): p. 542-550.
• Referans46. Pichery, M. et al.,(2012). Endogenous IL-33 is highly expressed in mouse epithelial barrier tissues, lymphoid organs, brain, embryos, and inflamed tissues: in situ analysis using a novel Il-33– LacZ gene trap reporter strain. The Journal of Immunology. 188 (7): p. 3488-3495.
• Referans47. Randow, F. et al., (1995). Mechanism of endotoxin desensitization: involvement of interleukin 10 and transforming growth factor beta. The Journal of experimental medicine. 181 (5): p. 1887- 1892.
• Referans48. Shapouri-Moghaddam A. et al.,(2018). Macrophage plasticity, polarization, and function in health and disease. Journal of cellular physiology. 233 (9): p. 6425-6440.
• Referans49. Sharma, R. and Anker S.D ,(2002). Cytokines, apoptosis and cachexia: the potential for TNF antagonism. International Journal of Cardiology. 85 (1): p. 161-171.
• Referans50. Schmitz, J. et al.,(2005). 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. 23 (5): p. 479-490.
• Referans51. Schroder, K. et al.,(2004). Interferon-γ: an overview of signals, mechanisms and functions. Journal of leukocyte biology. 75 (2): p. 163-189.
• Referans52. Schwartz, D.M. et al.,(2016). Type I/II cytokines, JAKs, and new strategies for treating autoimmune diseases. Nature Reviews Rheumatology. 12 (1): p. 25-36.
• Referans53. Tall, A.R. and Yvan-Charvet L,(2015). Cholesterol, inflammation and innate immunity. Nature Reviews Immunology. 15 (2): p. 104-116.
• Referans54. Tominaga, S.İ., (1989). A putative protein of a growth specific cDNA from BALB/c-3T3 cells is highly similar to the extracellular portion of mouse interleukin 1 receptor. FEBS letters. 258 (2): p. 301-304.
• Referans55.Villarino, A.V.,Kanno Y. and O'Shea J.J.,(2017). Mechanisms and consequences of Jak–STAT signaling in the immune system. Nature immunology. 18 (4): p. 374-384.
• Referans56. Viola, J. and Soehnlein O, (2015). Atherosclerosis–a matter of unresolved inflammation . in Seminars in immunology. Elsevier.
• Referans57. Weber, A., Wasiliew P. and Kracht, M.,(2010). Interleukin-1 (IL-1) pathway. Science signaling. 3 (105): p. cm1-cm1.
• Referans58. Wynn, T.A., Chawla, A. and Pollard, J.W.,(2013). Macrophage biology in development, homeostasis and disease. Nature. 496 (7446): p. 445-455.
• Referans59. Xin, P. et al.,(2020). The role of JAK/STAT signaling pathway and its inhibitors in diseases. International Immunopharmacology. 80 : p. 106-210.
• Referans60. Xu, H. et al.,(2019). Deficiency in IL-33/ST2 Axis Reshapes Mitochondrial Metabolism in Lipopolysaccharide-Stimulated Macrophages. Frontiers in Immunology. 10 .
• Referans61. Yang, J. et al.,(2014). Monocyte and macrophage differentiation: circulation inflammatory monocyte as biomarker for inflammatory diseases. Biomarker research. 2(1): p. 1-9.
• Referans62. Yu, H., Pardoll, D. and Jove, R.,(2009). STATs in cancer inflammation and immunity: a leading role for STAT3. Nature reviews cancer. 9 (11): p. 798-809.
• Referans63. Yue, S. et al.,(2014). Myeloid PTEN deficiency protects livers from ischemia reperfusion injury by facilitating M2 macrophage differentiation. The Journal of Immunology. 192 (11): p. 5343- 5353.
• Referans64. Zhang, X. et al.,(2019), Active vitamin D regulates macrophage M1/M2 phenotypes via the STAT-1- TREM-1 pathway in diabetic nephropathy. Journal of cellular physiology. 234 (5): p. 6917-6926.