Background. Regulatory T (Treg) cell plays a key role in autoimmune diseases. We evaluated the regulatory function and frequency of Treg cells and secreted IL-10, IL-35 concentration in Crohn’s disease (CD).
Methods. Twenty-three patients with CD and 25 healthy controls (HC) were included in this study. We analysed the alteration of Tregs frequency using flow cytometry for CD4, CD25, CD127 and FoxP3 markers. Surface expression of CD4, CD25 and CD127 markers were used for isolation of relatively pure Treg cells. Suppressive activity of Tregs was determined by measuring their ability to inhibit the proliferation of T responder (Tres) cells. In addition, the amounts of IL-10 and IL-35 cytokines in co-culture supernatants were measured by ELISA assay after stimulation with anti-CD2/CD3/CD28.
Results. CD patients had significantly lower frequency of CD4+ CD25+ CD127low FoxP3+ Treg cells in comparison with controls (2.17 ± 1.04 vs. 2.83 ± 1.07, p = 0.0352). Additionally, Treg cells mediated suppression was not significantly different in CD patients compared to controls. There was a significant difference in IL-10 secretion in response to anti-CD2/CD3/CD28 stimulation compared with HC (p = 0.0074).
Conclusion. The frequency of CD4+ CD25+ CD127low FoxP3+ Tregs decreased in active stage of CD but there was no impaired suppressive function of CD4+ CD25+ CD127low FoxP3+ Treg cells. We suggest that an alteration in the balance of Tregs and T effectors may contribute to pathogenesis of CD.
1. FLOYD DN, LANGHAM S, SÉVERAC HC, LEVESQUE BG. The economic and quality-of-life burden of Crohn’s disease in Europe and the United States, 2000 to 2013: a systematic review. Digestive diseases and sciences. 2015; 60(2):299-312.
2. EBRAHIMPOUR S, SHAHBAZIM, KHALILI A, TAHOORI MT, ZAVARAN HOSSEINI A, AMARI A, et al. Elevated levels of IL-2 and IL-21 produced by CD4+ T cells in inflammatory bowel disease. J Biol Regul Homeost Agents. 2017; 31(2):279-87.
3. HASANJANI RM, BAYANI M, SOLEIMANI AS, MOHAMMADNIA-AFROUZI M, NOURI H, EBRAHIMPOUR S. Evaluation of CD4+ CD25+ FoxP3+ regulatory T cells during treatment of patients with brucellosis. Journal of biological regulators and homeostatic agents. 2016; 30(3):675.
4. GRANT CR, LIBERAL R, MIELI-VERGANI G, VERGANI D, LONGHI MS. Regulatory T-cells in autoimmune diseases: challenges, controversies and—yet—unanswered questions. Autoimmunity reviews. 2015; 14(2):105-16.
5. MOHAMMADNIA-AFROUZI M, ZAVARAN HOSSEINI A, KHALILI A, ABEDIANKENARI S, HOSSEINI V, MALEKI I. Decrease of CD4+ CD25+ CD127low FoxP3+ regulatory T cells with impaired suppressive function in untreated ulcerative colitis patients. Autoimmunity. 2015; 48(8):556-61.
6. MOHAMMADNIA-AFROUZI M, SHAHBAZI M, DAMAVANDI SB, GANJI GF, EBRAHIMPOUR S. Regulatory T-cell: Regulator of Host Defense in Infection. Journal of Molecular Biology Research. 2017; 7(1):9.
7. BACCHETTA R, BARZAGHI F, RONCAROLO MG. From IPEX syndrome to FOXP3 mutation: a lesson on immune dysregulation. Annals of the New York Academy of Sciences. 2016.
8. CAUDY AA, REDDY ST, CHATILA T, ATKINSON JP, VERBSKY JW. CD25 deficiency causes an immune dysregulation, polyendocrinopathy, enteropathy, X-linked–like syndrome, and defective IL-10 expression from CD4 lymphocytes. Journal of Allergy and Clinical Immunology. 2007; 119(2):482-7.
9. LI Y, WANG Y, LIU Y, WANG Y, ZUO X, LI Y, et al. The Possible Role of the Novel Cytokines IL-35 and IL-37 in Inflammatory Bowel Disease. Mediators of Inflammation. 2014; 2014:10.
10. LIU Z, FENG B-S, YANG S-B, CHEN X, SU J, YANG P-C. Interleukin (IL)-23 suppresses IL-10 in inflammatory bowel disease. Journal of Biological Chemistry. 2012; 287(5):3591-7.
11. HARVEY RF, BRADSHAW JM. A simple index of Crohn’s-disease activity. Lancet. 1980; 1(8167):514.
12. KONTIAINEN S, SCHEININT, HALME L. Number of activated T-helper cells and NK cells in peripheral blood is decreased in severe Crohn’s disease. Apmis. 1996; 104(5):355-61.
13. DI SABATINO A, BIANCHERI P, PICONESE S, ROSADO MM, ARDIZZONE S, ROVEDATTI L, et al. Peripheral regulatory T cells and serum transforming growth factor-β: Relationship with clinical response to infliximab in Crohn’s disease. Inflammatory bowel diseases. 2010; 16(11):1891-7.
14. MAUL J, LODDENKEMPER C, MUNDT P, BERG E, GIESE T, STALLMACH A, et al. Peripheraland intestinal regulatory CD4+ CD25 high T cells in inflammatory bowel disease. Gastroenterology. 2005; 128(7):1868-78.
15. GUIDI L, FELICE C, PROCOLI A, BONANNO G, MARTINELLI E, MARZO M, et al. FOXP3(+) T Regulatory Cell Modifications in Inflammatory Bowel Disease Patients Treated with Anti-TNFα Agents. BioMed Research International. 2013; 2013:286368.
16. MAKITA S, KANAI T, OSHIMA S, URAUSHIHARA K, TOTSUKA T, SAWADA T, et al. CD4+ CD25bright T cells in human intestinal lamina propria as regulatory cells. The Journal of Immunology. 2004; 173(5):3119-30.
17. CUADRADO E, ALONSO M, DE JUAN MD, ECHANIZ P, ARENAS JI. Regulatory T cells in patients with inflammatory bowel diseases treated with adacolumn granulocytapheresis. World J Gastroenterol. 2008; 14(10):15217.
18. UENO A, JIJON H, CHAN R, FORD K, HIROTA C, KAPLAN GG, et al. Increased prevalence of circulating novel IL-17 secreting Foxp3 expressing CD4+ T cells and defective suppressive function of circulating Foxp3+ regulatory cells support plasticity between Th17 and regulatory T cells in inflammatory bowel disease patients. Inflamm Bowel Dis. 2013; 19(12):2522-34.
19. SARUTA M, YU QT, FLESHNER PR, MANTEL PY, SCHMIDT-WEBER CB, BANHAM AH, et al. Characterization of FOXP3+CD4+ regulatory T cells in Crohn’s disease. Clin Immunol. 2007;125(3):281-90.
20. Sun M, He C, Cong Y, Liu Z. Regulatory immune cells in regulation of intestinal inflammatory response to microbiota. Mucosal immunology. 2015; 8(5):969-78.
21. CHAUDHRY A, SAMSTEIN RM, TREUTING P, LIANG Y, PILSMC, HEINRICH J-M, et al. Interleukin-10 signaling in regulatory T cells is required for suppression of Th17 cell-mediated inflammation. Immunity. 2011; 34(4):566-78.
22. COLLISON LW, CHATURVEDI V, HENDERSON AL, GIACOMIN PR, GUY C, BANKOTI J, et al. IL-35-mediated induction of a potent regulatory T cell population. Nature immunology. 2010; 11(12):1093-101.
23. FRANKE A, BALSCHUN T, KARLSEN TH, SVENTORAITYTE J, NIKOLAUS S, MAYR G, et al. Sequence variants in IL10, ARPC2 and multiple other loci contribute to ulcerative colitis susceptibility. Nat Genet. 2008; 40(11):1319-23.
24. IYER SS, CHENG G. Role of Interleukin 10 Transcriptional Regulation in Inflammation and Autoimmune Disease. Critical reviews in immunology. 2012; 32(1):23-63.
25. MOHAMMADNIA-AFROUZI M, HOSSEINI AZ, KHALILI A, ABEDIANKENARI S, AMARI A, AGHILI B, et al. Altered microRNA expression and immunosuppressive cytokine production by regulatory T cells of ulcerative colitis patients. Immunological investigations. 2016; 45(1):63-74.