Original Article

The Impact of Vitamin D Supplementation on the IFNγ-IP10 Axis in Women with Hashimoto’s Thyroiditis Treated with Levothyroxine: A Double-blind Randomized Placebo-controlled Trial


Hashimoto's thyroiditis (HT) results from chemoattraction of inflammatory cells toward the thyroid gland by inducing the production of interferon-gamma (IFNγ)-induced protein 10 (IP10) by T helper (Th) 1 cells. Vitamin D may suppress the IFNγ-IP10 axis, but this new function of vitamin D has not yet been investigated in HT patients.
In an intervention and control group, patients received 50000 IU cholecalciferol or placebo every week for three months, respectively. The CD4+ T cells of 40 patients were isolated, and the mRNA expression levels of vitamin D receptor (VDR), peroxisome proliferator-activated receptors (PPAR)-α, and PPAR-γ genes were determined by real-time PCR. ELISA method was used to determine serum levels of vitamin D, tumor necrosis factor-alpha (TNF-α), IFN-γ, and IP10.
Vitamin D levels in the intervention group were significantly higher than in the placebo group after supplementation. PPAR-α and PPAR-γ gene expression levels did not differ significantly between the two groups. The serum levels of IP10, IFNγ, and TNF-α decreased significantly in the vitamin D group, as well as in the placebo group. 
During this study, vitamin D levels significantly increased in the intervention group and inflammatory factors decreased. Based on the similar results obtained in the placebo group, further studies with larger sample sizes and longer intervention times are recommended.

1. Tozzoli R, Barzilai O, Ram M, Villalta D, Bizzaro N, Sherer Y, et al. Infections and autoimmune thyroid diseases: parallel detection of antibodies against pathogens with proteomic technology. Autoimmun Rev. 2008;8(2):112-5.
2. García-López MA, Sancho D, Sánchez-Madrid F, Marazuela M. Thyrocytes from autoimmune thyroid disorders produce the chemokines IP-10 and Mig and attract CXCR3+ lymphocytes. J Clin Endocrinol Metab. 2001;86(10):5008-16.
3. Burek CL, Rose NR. Autoimmune thyroiditis and ROS. Autoimmun Rev. 2008;7(7):530-7.
4. McLachlan SM, Rapoport B. Breaking tolerance to thyroid antigens: changing concepts in thyroid autoimmunity. Endocr Rev. 2014;35(1):59-105.
5. Guarneri F, Benvenga S. Environmental factors and genetic background that interact to cause autoimmune thyroid disease. Curr Opin Endocrinol Diabetes Obes. 2007;14(5):398-409.
6. McLeod DS, Cooper DS. The incidence and prevalence of thyroid autoimmunity. Endocrine. 2012;42(2):252-65.
7. Antonelli A, Ferrari SM, Frascerra S, Galetta F, Franzoni F, Corrado A, et al. Circulating chemokine (CXC motif) ligand (CXCL) 9 is increased in aggressive chronic autoimmune thyroiditis, in association with CXCL10. Cytokine. 2011;55(2):288-93.
8. Rotondi M, Chiovato L. The chemokine system as a therapeutic target in autoimmune thyroid diseases: a focus on the interferon-γ inducible chemokines and their receptor. Curr Pharm Des. 2011;17(29):3202-16.
9. Antonelli A, Ferrari SM, Giuggioli D, Ferrannini E, Ferri C, Fallahi P. Chemokine (C-X-C motif) ligand (CXCL)10 in autoimmune diseases. Autoimmun Rev. 2014;13(3):272-80.
10. Doron H, Amer M, Ershaid N, Blazquez R, Shani O, Lahav TG, et al. Inflammatory Activation of Astrocytes Facilitates Melanoma Brain Tropism via the CXCL10-CXCR3 Signaling Axis. Cell Rep. 2019;28(7):1785-98. e6.
11. Oelkrug C, Ramage J. Enhancement of T cell recruitment and infiltration into tumours. Clin Exp Immunol. 2014;178(1):1-8.
12. Kemp EH, Metcalfe RA, Smith KA, Woodroofe MN, Watson PF, Weetman AP. Detection and localization of chemokine gene expression in autoimmune thyroid disease. Clin Endocrinol (Oxf). 2003;59(2):207-13.
13. Antonelli A, Rotondi M, Ferrari SM, Fallahi P, Romagnani P, Franceschini SS, et al. Interferon-γ-inducible α-chemokine CXCL10 involvement in Graves’ ophthalmopathy: modulation by peroxisome proliferator-activated receptor-γ agonists. J Clin Endocrinol Metab. 2006;91(2):614-20.
14. D'Ambrosio D, Cippitelli M, Cocciolo MG, Mazzeo D, Di Lucia P, Lang R, et al. Inhibition of IL-12 production by 1, 25-dihydroxyvitamin D3. Involvement of NF-kappaB downregulation in transcriptional repression of the p40 gene. J Clin Invest. 1998;101(1):252-62.
15. Harant H, Wolff B, Lindley IJ. 1α, 25‐Dihydroxyvitamin D3 decreases DNA binding of nuclear factor‐κB in human fibroblasts. FEBS letters. 1998;436(3):329-34.
16. Wu S, Liao AP, Xia Y, Li YC, Li J-D, Sartor RB, et al. Vitamin D receptor negatively regulates bacterial-stimulated NF-κB activity in intestine. Am J Pathol. 2010;177(2):686-97.
17. Wu S, Xia Y, Liu X, Sun J. Vitamin D receptor deletion leads to reduced level of IκBα protein through protein translation, protein–protein interaction, and post-translational modification. Int J Biochem Cell Biol. 2010;42(2):329-36.
18. Yu X-P, Bellido T, Manolagas SC. Down-regulation of NF-kappa B protein levels in activated human lymphocytes by 1, 25-dihydroxyvitamin D3. Proc Natl Acad Sci U S A. 1995;92(24):10990-4.
19. Mayne CG, Spanier JA, Relland LM, Williams CB, Hayes CE. 1, 25‐Dihydroxyvitamin D3 acts directly on the T lymphocyte vitamin D receptor to inhibit experimental autoimmune encephalomyelitis. Eur J Immunol. 2011;41(3):822-32.
20. Grishkan IV, Fairchild AN, Calabresi PA, Gocke AR. 1, 25-Dihydroxyvitamin D3 selectively and reversibly impairs T helper-cell CNS localization. Proc Natl Acad Sci U S A. 2013;110(52):21101-6.
21. Harris SG, Phipps RP. The nuclear receptor PPAR gamma is expressed by mouse T lymphocytes and PPAR gamma agonists induce apoptosis. Eur J Immunol. 2001;31(4):1098-105.
22. Jones DC, Ding X, Daynes RA. Nuclear receptor PPARα is expressed in resting murine lymphocytes: the PPARα in T and B lymphocytes is both transactivation and transrepression competent. J Biol Chem. 2001.
23. Padilla J, Kaur K, Cao HJ, Smith TJ, Phipps RP. Peroxisome Proliferator Activator Receptor-γ Agonists and 15-Deoxy-Δ12, 1412, 14-PGJ2 Induce Apoptosis in Normal and Malignant B-Lineage Cells. J Immunol. 2000;165(12):6941-8.
24. Gosset P, Charbonnier AS, Delerive P, Fontaine J, Staels B, Pestel J, et al. Peroxisome proliferator‐activated receptor γ activators affect the maturation of human monocyte‐derived dendritic cells. Eur J Immunol. 2001;31(10):2857-65.
25. Marx N, Mach F, Sauty A, Leung JH, Sarafi MN, Ransohoff RM, et al. Peroxisome proliferator-activated receptor-γ activators inhibit IFN-γ-induced expression of the T cell-active CXC chemokines IP-10, Mig, and I-TAC in human endothelial cells. J Immunol. 2000;164(12):6503-8.
26. Schaefer KL, Denevich S, Ma C, Cooley SR, Nakajima A, Wada K, et al. Intestinal antiinflammatory effects of thiazolidenedione peroxisome proliferator-activated receptor-γ ligands on T helper type 1 chemokine regulation include nontranscriptional control mechanisms. Inflamm Bowel Dis. 2005;11(3):244-52.
27. Sertznig P, Seifert M, Tilgen W, Reichrath J. Activation of vitamin D receptor (VDR)- and peroxisome proliferator-activated receptor (PPAR)-signaling pathways through 1,25(OH)(2)D(3) in melanoma cell lines and other skin-derived cell lines. Dermatoendocrinol. 2009;1(4):232-8.
28. Antonelli A, Rotondi M, Fallahi P, Romagnani P, Ferrari SM, Buonamano A, et al. High levels of circulating CXC chemokine ligand 10 are associated with chronic autoimmune thyroiditis and hypothyroidism. J Clin Endocrinol Metab. 2004;89(11):5496-9.
29. Rong L, Li K, Li R, Liu HM, Sun R, Liu XY. Analysis of tumor-infiltrating gamma delta T cells in rectal cancer. World J Gastroenterol. 2016;22(13):3573-80.
30. Kivity S, Agmon-Levin N, Zisappl M, Shapira Y, Nagy EV, Dankó K, et al. Vitamin D and autoimmune thyroid diseases. Cell Mol Immunol. 2011;8(3):243-7.
31. Chailurkit L-o, Aekplakorn W, Ongphiphadhanakul B. High vitamin D status in younger individuals is associated with low circulating thyrotropin. Thyroid. 2013;23(1):25-30.
32. Choi YM, Kim WG, Kim TY, Bae SJ, Kim H-K, Jang EK, et al. Low levels of serum vitamin D3 are associated with autoimmune thyroid disease in pre-menopausal women. Thyroid. 2014;24(4):655-61.
33. Zhang Q, Wang Z, Sun M, Cao M, Zhu Z, Fu Q, et al. Association of high vitamin d status with low circulating thyroid-stimulating hormone independent of thyroid hormone levels in middle-aged and elderly males. Int J Endocrinol. 2014;2014.
34. Mackawy AMH, Al-Ayed BM, Al-Rashidi BM. Vitamin D deficiency and its association with thyroid disease. IJHSP. 2013;7(3):267.
35. Charo IF, Ransohoff RM. The many roles of chemokines and chemokine receptors in inflammation. N Engl J Med. 2006;354(6):610-21.
36. Xie JH, Nomura N, Lu M, Chen SL, Koch GE, Weng Y, et al. Antibody-mediated blockade of the CXCR3 chemokine receptor results in diminished recruitment of T helper 1 cells into sites of inflammation. J Leukoc Biol. 2003;73(6):771-80.
37. Campanella GS, Medoff BD, Manice LA, Colvin RA, Luster AD. Development of a novel chemokine-mediated in vivo T cell recruitment assay. J Immunol Methods. 2008;331(1-2):127-39.
38. Romagnani P, Maggi L, Mazzinghi B, Cosmi L, Lasagni L, Liotta F, et al. CXCR3-mediated opposite effects of CXCL10 and CXCL4 on TH1 or TH2 cytokine production. J Allergy Clin Immunol. 2005;116(6):1372-9.
39. Rotondi M, Chiovato L, Romagnani S, Serio M, Romagnani P. Role of chemokines in endocrine autoimmune diseases. Endocr Rev. 2007;28(5):492-520.
40. Hayes CE, Hubler SL, Moore JR, Barta LE, Praska CE, Nashold FE. Vitamin D Actions on CD4(+) T Cells in Autoimmune Disease. Frontiers in immunology. 2015;6:100.
41. Antonelli A, Rotondi M, Fallahi P, Romagnani P, Ferrari SM, Buonamano A, et al. High levels of circulating CXC chemokine ligand 10 are associated with chronic autoimmune thyroiditis and hypothyroidism. J Clin Endocrinol Metab. 2004;89(11):5496-9.
42. Liu C, Papewalis C, Domberg J, Scherbaum WA, Schott M. Chemokines and autoimmune thyroid diseases. Horm Metab Res. 2008;40(6):361-8.
43. Corona G, Biagini C, Rotondi M, Bonamano A, Cremonini N, Petrone L, et al. Correlation between, clinical, biochemical, color Doppler ultrasound thyroid parameters, and CXCL-10 in autoimmune thyroid diseases. Endocr J. 2008;55(2):345-50.
44. Antonelli A, Fallahi P, Rotondi M, Ferrari SM, Romagnani P, Grosso M, et al. Increased serum CXCL10 in Graves’ disease or autoimmune thyroiditis is not associated with hyper-or hypothyroidism per se, but is specifically sustained by the autoimmune, inflammatory process. Eur J Endocrinol. 2006;154(5):651-8.
45. Caturegli P, Hejazi M, Suzuki K, Dohan O, Carrasco N, Kohn LD, et al. Hypothyroidism in transgenic mice expressing IFN-gamma in the thyroid. Proc Natl Acad Sci U S A. 2000;97(4):1719-24.
46. Baud V, Karin M. Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol. 2001;11(9):372-7.
47. Zelová H, Hošek J. TNF-α signalling and inflammation: interactions between old acquaintances. Inflamm Res. 2013;62(7):641-51.
48. Zhang N, Wang Q, Tian Y, Xiong S, Li G, Xu L. Expressions of IL-17 and TNF-α in patients with Hashimoto's disease combined with thyroid cancer before and after surgery and their relationship with prognosis. Clin Transl Oncol. 2020;22(8):1280-7.
49. Botelho IMB, Moura Neto A, Silva CA, Tambascia MA, Alegre SM, Zantut-Wittmann DE. Vitamin D in Hashimoto's thyroiditis and its relationship with thyroid function and inflammatory status. Endocr J. 2018;65(10):1029-37.
50. Taheriniya S, Arab A, Hadi A, Fadel A, Askari G. Vitamin D and thyroid disorders: a systematic review and Meta-analysis of observational studies. BMC Endocr Disord. 2021;21(1):171.
51. Krysiak R, Szkróbka W, Okopień B. The Effect of Vitamin D on Thyroid Autoimmunity in Levothyroxine-Treated Women with Hashimoto's Thyroiditis and Normal Vitamin D Status. Exp Clin Endocrinol Diabetes. 2017;125(4):229-33.
52. Kim D. The Role of Vitamin D in Thyroid Diseases. Int J Mol Sci. 2017;18(9).
53. Kong N, Lan Q, Su W, Chen M, Wang J, Yang Z, et al. Induced T regulatory cells suppress osteoclastogenesis and bone erosion in collagen-induced arthritis better than natural T regulatory cells. Ann Rheum Dis. 2012;71(9):1567-72.
54. Zhao R, Zhang W, Ma C, Zhao Y, Xiong R, Wang H, et al. Immunomodulatory Function of Vitamin D and Its Role in Autoimmune Thyroid Disease. Frontiers in immunology. 2021;12:574967.
55. Nodehi M, Ajami A, Izad M, Asgarian Omran H, Chahardoli R, Amouzegar A, et al. Effects of vitamin D supplements on frequency of CD4(+) T-cell subsets in women with Hashimoto's thyroiditis: a double-blind placebo-controlled study. Eur J Clin Nutr. 2019;73(9):1236-43.
56. Liontiris MI, Mazokopakis EE. A concise review of Hashimoto thyroiditis (HT) and the importance of iodine, selenium, vitamin D and gluten on the autoimmunity and dietary management of HT patients.Points that need more investigation. Hell J Nucl Med. 2017;20(1):51-6.
57. Zhang J, Chen Y, Li H, Li H. Effects of vitamin D on thyroid autoimmunity markers in Hashimoto's thyroiditis: systematic review and meta-analysis. J Int Med Res. 2021;49(12):3000605211060675.
IssueVol 21 No 4 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijaai.v21i4.10288
CD4-positive T-lymphocytes Hashimoto disease Interferon-gamma Peroxisome proliferator-activated receptors Th1 cells Vitamin D

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Robat-Jazi B, Mobini S, Chahardoli R, Mansouri F, Nodehi M, Esfahanian F, Saboor Yaraghi AA. The Impact of Vitamin D Supplementation on the IFNγ-IP10 Axis in Women with Hashimoto’s Thyroiditis Treated with Levothyroxine: A Double-blind Randomized Placebo-controlled Trial. Iran J Allergy Asthma Immunol. 2022;21(4):407-417.