Effects of 1,25-dihydroxyvitamin D3 on IL-17/IL-23 axis, IFN-γ and IL-4 expression in systemic lupus erythematosus induced mice model

Document Type: Original Article


1 Immunology Research Center, BuAli Research Institute, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pathology Department, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Systemic lupus erythematosus (SLE) is a multi-factorial autoimmune disease which may be characterized by T lymphocytes dysfunctions. Th17 cells have been identified as new effector cells, which play an important role in the pathogenesis. In recent years, immunomodulatory effect of vitamin D3 has been noticed. In the present experiment, the effect of vitamin D3 on the expression of IL-17, IL-23, IL-4 and IFN-γ were assessed in activated chromatin-induced mouse model for SLE.
Materials and Methods: Five groups of mice were included in this study; Group one received active chromatin +CFA + PBS; Group 2 received vitamin D3  starting 2 weeks before disease induction; Group 3 received vitamin D3 (50 ng/day) starting with the disease establishment; Group 4 received non active chromatin +CFA + PBS; Group 5 received CFA + PBS. On day 56 splenocytes were isolated and gene expression of interleukin IL-17, IL-23, IL-4 and IFN-γ were analyzed by Real-Time PCR method. Proteinuria and serum anti-dsDNA and Th17 levels were measured using commercial kits.
Results: The results showed that IL-17, IL-23, and IFN-γ mRNA expression, and IL-17 titers were decreased remarkably and that of IL-4 increased in mice which received vitamin D3 before SLE induction. Administration of vitamin D3 after the establishment of SLE failed to affect the IL-17 or IL-23 mRNA levels. Lastly, pre-treatment of mice with vitamin D3 decreased the anti-ds DNA antibody titer.
Conclusion: Our findings showed that vitamin D3 supplementation in lupus induced mice through modulating the expression rate of some inflammatory cytokines diminished the inflammatory conditions in SLE.


1. Tsokos GD. Systemic lupus erythematosus. New Engl J Med 2011; 365: 2110-2121.

2. D. P. D’Cruz, M. A. Khamashta, and G. R. Hughes, “Systemic lupus erythematosus.  Lancet 2007; 369: 587– 596.

3. Lewis JE, Fu SM, Gaskin F. Autoimmunity, end organ damage, and the origin of autoantibodies and autoreactive T cells in systemic lupus erythematosus. Discov Med 2013; 15: 85-92.

4. Perry D, Sang A, Yin Y, Zheng YY, Morel L. Murine models of systemic lupus erythematosus. J Biomed Biotech 2011; 2011: 271694.

5. Qiao B, Wu J, Chu YW, Wang Y, Wang DP, Wu HS et al.  Induction of systemic lupus erythematosus-like syndrome in syngeneic mice by immunization with activated lymphocyte-derived DNA. Rheumatology (Oxford) 2005; 44:1108-1114.

6. Shah K, Lee WW, Lee SH, Kim SH, Kang SW, Craft J et al. Dysregulated balance of TH17 and TH1 cells in systemic lupus erythematosus. Arthritis Res Ther 2010; 12: R53.

7. Basso AS, Cheroutre H, Mucida D. More stories on TH17 cells. Cell Res 2009; 19: 399-411.

8. Balanescu P, Balanescu E, Tanasescu C, Nicolau A, Tanasescu R, Grancea C, et al. TH17 cell population in lupus erythematosus. Rom J Intern Med 2010; 48: 255-259.

9. Apostolidis S A, Crisp´ın JC, Tsokos GC. IL-17- producing T cells in lupus nephritis. Lupus 2011; 20: 120–124.

10. Kleczynska W, Jakiela B, Plutecka H, Milewski M, Sanak M, Musial J. Imbalance between TH17 and regulatory T-cells in systemic lupus erythematosus. Folia Histochem Cytobiol 2011; 49: 646-653.

11. Manson JJ, Rahman A. Systemic lupus erythematosus. Orphanet. J Rare Dis 2006; 1: 6.

12. Ben-Menachem E. Review article: systemic lupus erythematosus: A review for anesthesiologists. Anesth Analg 2010; 111: 665-676.

13. Crispin JC, Liossis SN, Kis-Toth K, Kis-Toth K, Lieberman LA, Kyttaris VC, et al. Pathogenesis of human systemic lupus erythematosus: Recent advances. Trends Mol Med 2010; 16: 47-57.

14. Cutolo M. Further emergent evidence for the vitamin D endocrine system involvement in autoimmune rheumatic disease risk and prognosis. Ann Rheum Dis 2013; 72: 473-475.

15. Szodoray P, Nakken B, Gaal J, Jonsson R, Szegedi A, Zold E, et al. Complex role of vitamin D in autoimmune diseases. Scand J Immunol 2008; 68: 261-269.

16. Chang SH, Chung Y, Dong C. Vitamin D suppresses TH17 cytokine production by inducing C/EBP homologous protein (CHOP) expression. J Biol Chem 2010; 285: 38751-38755.

17. Wen ZK, Xu W, Xu L, Cao OH, Wang Y, Chu YW, et al.  DNA hypomethylation is crucial for apoptotic DNA to induce systemic lupus erythematosus-like autoimmune disease in SLE-nonsusceptible mice. Rheumatology 2007; 46: 1796-1803.

18. Apostolidis SA, Lieberman LA, Kis-Toth K, Crispin JC, Tsokos GC. The dysregulation of cytokine networks in systemic lupus erythematosus. J Interferon Cytokine Res 2011; 31: 769-779.

19. Ding-Lei Su ZM, Min-Ning S, Xia L, Ling-Yun S. Roles of pro- and anti-inflammatory cytokines in the pathogenesis of SLE. J Biomed Biotech 2012; 2012: 347141.

20. Edgerton C, Crispín JC, Moratz CM, Betteli E, Oukka M, Simovic M, et al. IL-17 producing CD4+ T-cells mediate accelerated ischemia/reperfusion-induced injury in autoimmunity-prone mice. Clin Immunol 2009; 130: 313-321.

21. Zhang Z, Kyttaris VC, Tsokos GC. The role of IL-23/IL-17 axis in lupus nephritis. J Immunol 2009; 183: 3160-3169.

22. Kyttaris VC, Zhang Z, Kuchroo VK, Oukka M, Tsokos GC. Cutting Edge: IL-23 receptor deficiency orevents development of lupus nephritis in C57BL/6–lpr/lpr mice. J Immunol 2010; 184: 4605-4609.

23. Tabasi N, Rastin M, Mahmoudi M, Ghoryani M, Mirfeizi Z, Zamani SH, et al. Influence of Vitamin D on cell cycle, apoptosis, and some apoptosis related molecules in systemic lupus erythematosus. Iran J Basic Med Sci 2015; 18: 1107-1111.

24. Lavi Arab F, Rastin M, Faraji F, Zamani Taghizadeh Rabe S, Tabasi N, Khazaee M, et al. Assessment of 1,25-D3 effects on Treg cells and their related molecules in active chromatin lupus induced Balb/c mice. Immunopharmacol Immunotoxicol 2015; 37: 12-18.

25. Wahono CS, Rusmini H, Soelistyoningsih D, Hakim R, Handono K, Endharti AT, et al. Effects of 1,25(OH)2D3 in immune response regulation of systemic lupus erithematosus (SLE) patient with hypovitamin D. Int J Clin Exp Med 2014; 7: 22-31.

26. Lemire JM, Ince A, Takashima M. 1,25-Dihydroxyvitamin D3 attenuates expression of experimental murine lupus in MRL/1 mice. Autoimmunity 1992; 12:143-148.

27. Deluca HF, Cantorna MT. Vitamin D: Its role and uses in immunology. FASEB J 2001; 15: 2579-2585.

28. Koizumi T, Nakao Y, Matsui T, Nakagawa T, Matsuda S, Komoriya K, et al. Effects of corticosteroid and 1,24R-dihydroxy-vitamin D3 administration on lymphoproliferation and autoimmune disease in MRL/MP-lpr/lpr mice. Int Arch Allergy Appl Immunol 1985; 77: 396-404.

29. Vaisberg MW, Kaneno R, Franco MF, Mendes NF. Influence of cholecalci-ferol (vitamin D3) on course of experimental systemic lupus erythematosus in F1 (NZB x W) mice. J Clin Lab Anal 2000; 14: 91-96.

30. Reihani H, Rastin M, Mahmoudi M, Ghoryani M, Abdlahi N, Tabasi N, et al. Influence of 1 Alpha, 25-Dihydroxyvitamin D3 on T Helper 17 Cells and Related Cytokines in Systemic Lupus

Erythematosus, Iran J Immunol 2015; 12: 82-93.

31. Onishi RM, Gaffen SL. Interleukin-17 and its target genes: mechanisms of interleukin-17 function in disease. Immunology 2010; 129:311–321.

32.  Langrish CL, Chen Y, Blumenschein WM, Mattson J, Basham B, Sedgwick JD, et al. IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. J Exp Med 2005; 201:233–240

33. Raza A, Yousaf W, Giannella R, Shata M. Th17 cells: interactions with predisposing factors in the immunopathogenesis of inflammatory bowel disease. Expert Rev Clin Immunol 2012; 8: 161–168.

34. Daniel C, Sartory NA, Zahn N, Radeke HH,            Stein JM. Immune modulatory treatment of trinitrobenzene sulfonic acid colitis with calcitriol is associated with a change of a T-helper (TH) 1/TH17 to a TH2 and regulatory T-cell profile. J Pharmacol Exp Ther 2008; 324: 23-33.

35. Ding-Lei Su ZM, Min-Ning S, Xia L, Ling-Yun S. Roles of pro- and anti-inflammatory cytokines in the pathogenesis of SLE. J Biomed Biotech 2012; 2012: 347141.

36. Jeffery LE, Burke F, Mura M, Zheng Y, Qureshi OS, Hewison M, et al. 1,25 Dihydroxyvitamin D3 and IL-2 combine to inhibit T Cell production of inflammatory cytokines and promote development of regulatory T Cells expressing CTLA-4 and Foxp3. J Immunol 2009; 183: 5458-5467.

37. Terrier B, Derian N, Schoindre Y, Charra W, Geri G, Zahr N, et al. Restoration of regulatory and effector T cell balance and B cell homeostasis in systemic lupus erythematosus patients through vitamin D supplementation. Arthritis Res Therap 2012; 14: R22.

38. Singh A, Kamen DL. Potential benefits of vita­min D for patients with systemic lupus erythe­matosus. Dermatoendocrinol 2012; 4: 146-51.

39. Ghoryani M, Sahebari M, Mahmoudi M, Abdollahi N, Reihani H, Zamani SH, et al. Immunomodulatory Vitamin D effects on regulatory T-cells and cytokines in an in vitro study on patients with systemic lupus erythematosus. Food and Agricultural Immunology 2016; 27: 377-387.

40. Funauchi M, Yu H, Sugiyama M, Ikoma S, Ohno M, Kinoshita K, et al.  Increased IL-4 production by NK T-cells in systemic lupus erythematosus. Clin Immunol 1999; 92: 197-202.

41. Dean GS, Anand A, Blofeld A, Isenberg DA, Lydyard PM. Characterization of CD3+CD4-CD8- (double negative) T-cells in patients with systemic lupus erythematosus: Production of IL-4. Lupus 2002; 11: 501-507.