Protective effect of interleukin-36 receptor antagonist on liver injury induced by concanavalin A in mice

Document Type : Original Article


1 Department of Immunology, Medical School of Ningbo University, Ningbo 315211, China

2 Department of Hepatology, HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo 315010, China


Objective(s): Interleukin-36 receptor antagonist (IL-36Ra) is a new member of the IL-1 family that exhibits anti-inflammatory activity in a variety of inflammatory and immune diseases. Our purpose was to determine the effect of IL-36Ra on liver injury in a mouse hepatitis model induced by concanavalin A (ConA).
Materials and Methods: Mice were treated with IL-36Ra DNA or pcDNA3.1 control plasmid using a hydrodynamic gene delivery approach.
Results: Our data reveal that treatment with IL-36Ra decreased liver inflammation and serum level of aminotransferases. Furthermore, IL-36Ra reduced ConA-induced pro-inflammatory cytokines (interferon-γ, tumor necrosis factor-α, and IL-17A) production when compared to control plasmid.
Conclusion: Our results demonstrated that IL-36Ra is a critical protector against ConA-induced liver injury.


1. Gresnigt MS, van de Veerdonk FL. Biology of IL-36 cytokines and their role in disease. Semin Immunol 2013; 25: 458-465.
2. Carrier Y, Ma HL, Ramon HE, Napierata L, Small C, O’Toole M, et al. Inter-regulation of Th17 cytokines and the IL-36 cytokines in vitro and in vivo: implications in psoriasis pathogenesis. J Invest Dermatol 2011; 131: 2428-2437.
3. Barton JL, Herbst R, Bosisio D, Higgins L, Nicklin MJ. A tissue specific IL-1 receptor antagonist homolog from the IL-1 cluster lacks IL-1, IL-1ra, IL-18 and IL-18 antagonist activities. Eur J Immunol 2000; 30: 3299-3308.
4. Smith DE, Renshaw BR, Ketchem RR, Kubin M, Garka KE, Sims JE. Four new members expand the interleukin-1 superfamily. J Biol Chem 2000; 275: 1169-1175.
5. Johnston A, Xing X, Guzman AM, Riblett M, Loyd CM, Ward NL, et al. IL-1F5, -F6, -F8, and -F9: a novel IL-1 family signaling system that is active in psoriasis and promotes keratinocyte antimicrobial peptide expression. J Immunol 2011; 186: 2613-2622.
6. Chustz RT, Nagarkar DR, Poposki JA, Favoreto S, Jr., Avila PC, Schleimer RP, et al. Regulation and function of the IL-1 family cytokine IL-1F9 in human bronchial epithelial cells. Am J Respir Cell Mol Biol 2011; 45: 145-153.
7. Karumbaiah L, Norman SE, Rajan NB, Anand S, Saxena T, Betancur M, et al. The upregulation of specific interleukin (IL) receptor antagonists and paradoxical enhancement of neuronal apoptosis due to electrode induced strain and brain micromotion. Biomaterials 2012; 33: 5983-5996.
8. Tazi-Ahnini R, Cox A, McDonagh AJ, Nicklin MJ, di Giovine FS, Timms JM, et al. Genetic analysis of the interleukin-1 receptor antagonist and its homologue IL-1L1 in alopecia areata: strong severity association and possible gene interaction. Eur J Immunogenet 2002; 29: 25-30.
9. Lamacchia C, Palmer G, Rodriguez E, Martin P, Vigne S, Seemayer CA, et al. The severity of experimental arthritis is independent of IL-36 receptor signaling. Arthritis Res Ther 2013; 15: R38.
10. Palomo J, Dietrich D, Martin P, Palmer G, Gabay C. The interleukin (IL)-1 cytokine family: Balance between agonists and antagonists in inflammatory diseases. Cytokine 2015; 76: 25-37.
11. Debets R, Timans JC, Homey B, Zurawski S, Sana TR, Lo S, et al. Two novel IL-1 family members, IL-1 delta and IL-1 epsilon, function as an antagonist and agonist of NF-kappa B activation through the orphan IL-1 receptor-related protein 2. J Immunol 2001; 167: 1440-1446.
12. Towne JE, Garka KE, Renshaw BR, Virca GD, Sims JE. Interleukin (IL)-1F6, IL-1F8, and IL-1F9 signal through IL-1Rrp2 and IL-1RAcP to activate the pathway leading to NF-kappaB and MAPKs. J Biol Chem 2004; 279: 13677-13688.
13. Mutamba S, Allison A, Mahida Y, Barrow P, Foster N. Expression of IL-1Rrp2 by human myelomonocytic cells is unique to DCs and facilitates DC maturation by IL-1F8 and IL-1F9. Eur J Immunol 2012; 42: 607-617.
14. Carbone M, Neuberger JM. Autoimmune liver disease, autoimmunity and liver transplantation. J Hepatol 2014; 60: 210-223.
15. Tiegs G, Hentschel J, Wendel A. A T cell-dependent experimental liver injury in mice inducible by concanavalin A. J Clin Invest 1992; 90: 196-203.
16. Odashima M, Otaka M, Jin M, Horikawa Y, Matsuhashi T, Ohba R, et al. A selective adenosine A2A receptor agonist, ATL-146e, prevents concanavalin A-induced acute liver injury in mice. Biochem Biophys Res Commun 2006; 347: 949-954.
17. Yin H, Cheng L, Agarwal C, Agarwal R, Ju C. Lactoferrin protects against concanavalin A-induced liver injury in mice. Liver Int 2010; 30: 623-632.
18. Scheiermann P, Bachmann M, Hardle L, Pleli T, Piiper A, Zwissler B, et al. Application of IL-36 receptor antagonist weakens CCL20 expression and impairs recovery in the late phase of murine acetaminophen-induced liver injury. Sci Rep 2015; 5: 8521.
19. Liu F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther 1999; 6: 1258-1266.
20. Cao L, Zou Y, Zhu J, Fan X, Li J. Ginsenoside Rg1 attenuates concanavalin A-induced hepatitis in mice through inhibition of cytokine secretion and lymphocyte infiltration. Mol Cell Biochem 2013; 380: 203-210.
21. Gantner F, Leist M, Lohse AW, Germann PG, Tiegs G. Concanavalin A-induced T-cell-mediated hepatic injury in mice: the role of tumor necrosis factor. Hepatology 1995; 21: 190-198.
22. Lamacchia C, Rodriguez E, Palmer G, Vesin C, Seemayer CA, Rubbia-Brandt L, et al. Mice deficient in hepatocyte-specific IL-1Ra show delayed resolution of concanavalin A-induced hepatitis. Eur J Immunol 2012; 42: 1294-1303.
23. Bulau AM, Fink M, Maucksch C, Kappler R, Mayr D, Wagner K, et al. In vivo expression of interleukin-37 reduces local and systemic inflammation in concanavalin A-induced hepatitis. ScientificWorldJournal 2011; 11: 2480-2490.
24. Yuan X, Li Y, Pan X, Peng X, Song G, Jiang W, et al. IL-38 alleviates concanavalin A-induced liver injury in mice. Int Immunopharmacol 2016; 40: 452-457.
25. Xu JF, Xiao H, Hu GY, Zheng SH, Liu W, Yuan CL, et al. Ectopic B7-H4-Ig expression attenuates concanavalin A-induced hepatic injury. Clin Immunol 2010; 136: 30-41.
26. Xu X, Hu Y, Zhai X, Lin M, Chen Z, Tian X, et al. Salvianolic acid A preconditioning confers protection against concanavalin A-induced liver injury through SIRT1-mediated repression of p66shc in mice. Toxicol Appl Pharmacol 2013; 273: 68-76.
27. Zhou Y, Dai W, Lin C, Wang F, He L, Shen M, et al. Protective effects of necrostatin-1 against concanavalin A-induced acute hepatic injury in mice. Mediators Inflamm 2013; 2013: 706156.
28. Kremer M, Perry AW, Milton RJ, Rippe RA, Wheeler MD, Hines IN. Pivotal role of Smad3 in a mouse model of T cell-mediated hepatitis. Hepatology 2008; 47: 113-126.
29. Kusters S, Gantner F, Kunstle G, Tiegs G. Interferon gamma plays a critical role in T cell-dependent liver injury in mice initiated by concanavalin A. Gastroenterology 1996; 111: 462-471.
30. An MM, Fan KX, Cao YB, Shen H, Zhang JD, Lu L, et al. Lymphtoxin beta receptor-Ig protects from T-cell-mediated liver injury in mice through blocking LIGHT/HVEM signaling. Biol Pharm Bull 2006; 29: 2025-2030.
31. Tacke F, Luedde T, Trautwein C. Inflammatory pathways in liver homeostasis and liver injury. Clin Rev Allergy Immunol 2009; 36: 4-12.
32. Xu S, Han Y, Xu X, Bao Y, Zhang M, Cao X. IL-17A-producing gammadeltaT cells promote CTL responses against Listeria monocytogenes infection by enhancing dendritic cell cross-presentation. J Immunol 2010; 185: 5879-5887.
33. Zhao N, Hao J, Ni Y, Luo W, Liang R, Cao G, et al. Vgamma4 gammadelta T cell-derived IL-17A negatively regulates NKT cell function in Con A-induced fulminant hepatitis. J Immunol 2011; 187: 5007-5014.
34. Stout-Delgado HW, Du W, Shirali AC, Booth CJ, Goldstein DR. Aging promotes neutrophil-induced mortality by augmenting IL-17 production during viral infection. Cell Host Microbe 2009; 6: 446-456.
35. Kelly MN, Kolls JK, Happel K, Schwartzman JD, Schwarzenberger P, Combe C, et al. Interleukin-17/interleukin-17 receptor-mediated signaling is important for generation of an optimal polymorphonuclear response against Toxoplasma gondii infection. Infect Immun 2005; 73: 617-621.
36. Vigne S, Palmer G, Lamacchia C, Martin P, Talabot-Ayer D, Rodriguez E, et al. IL-36R ligands are potent regulators of dendritic and T cells. Blood 2011; 118: 5813-5823.
37. Vigne S, Palmer G, Martin P, Lamacchia C, Strebel D, Rodriguez E, et al. IL-36 signaling amplifies Th1 responses by enhancing proliferation and Th1 polarization of naive CD4+ T cells. Blood 2012; 120: 3478-3487.
38. Gresnigt MS, Rosler B, Jacobs CW, Becker KL, Joosten LA, van der Meer JW, et al. The IL-36 receptor pathway regulates Aspergillus fumigatus-induced Th1 and Th17 responses. Eur J Immunol 2013; 43: 416-426.