Numerical status of CD4+CD25+FoxP3+ and CD8+CD28- regulatory T cells in multiple sclerosis

Document Type : Original Article


1 Department of Neurology, Kashan University of Medical Sciences, Kashan, Iran

2 Student Research Committee, Kashan University of Medical Sciences, Kashan, Iran

3 Department of Pediatrics, Kashan University of Medical Sciences, Kashan, Iran

4 Nephrology and Urology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

5 Trauma Research center, Kashan University of Medical Sciences, Kashan, Iran


Objective(s): Regulatory T cells, including CD4+CD25+Fox3+ and CD8+CD28- cells play an important role in regulating the balance between immunity and tolerance. Since multiple sclerosis is an inflammatory autoimmune disease, regulatory T cells are considered to be involved in its pathogenesis. In this study, we investigated the circulatory numbers of the two mentioned types of regulatory T cells and also their association with different clinical characteristics in 84 multiple sclerosis patients.
Materials and Methods: 84 patients with multiple sclerosis and 75 normal individuals were studied. Demographic and clinical information of all participants were collected via questionnaire and clinical examination as well as MRI. The peripheral blood frequency of two different subgroups of regulatory T cells (CD4+ CD25+Foxp3+ and CD8+CD28- cells) were analyzed by flow cytometry using anti-human antibodies conjugated with CD4-FITC / CD25-PE/Foxp3-PE-Cy5, CD3-PE/CD8a-PE-Cy5/CD28-FITC.
Results: The frequency of CD4+CD25+Foxp3+ cells in multiple sclerosis patients was significantly less than that in healthy controls (P=0.006) and in mild forms less than that in sever forms (P=0.003). There was not any correlation between the frequency of regulatory T cells and different clinical variables.
Conclusion: Our results showed that the number of CD4+CD25+Foxp3+ cells decreases significantly in multiple sclerosis patients, which probably shows the regulatory role of these cells in multiple sclerosis.


1. Compston A, Coles A. Multiple sclerosis. Lancet 2008; 372:1502-1517.
2. Trapp B, Peterson J, Ransohoff R, Rudick R, Mork S, Bo L. Axonal transection in the lesions of multiple sclerosis. N Engl J Med 1998; 338:278–285.
3. Hafler DA, Slavik JM, Anderson DE, O'Connor KC, De Jager P, Baecher-AllanC. Multiple sclerosis. Immunol Rev 2005; 204:208–231.
4. Sakaguchi S, Sakaguchi N, Asano M, Itoh M, Toda M. Immunologic selftolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol 1995; 155:1151–1164.
5. Fritzsching B, Korporal M, Haas J, Krammer PH, Suri-Payer E, Wildemann B. Similar sensitivity of regulatory T cells toward CD95Lmediated apoptosis in patients with multiple sclerosis and healthy individuals.  J Neurol Sci 2006; 251:91–97.
6. Kukreja A, Cost G, Marker J,  Zhang C, Sun Z, Lin-Su K, et al. Multiple immuno-regulatory defects in type-1 diabetes. J Clin Invest  2002; 109:131–140.
7. Crucian B, Dunne P, Friedman H, Ragsdale R, Pross S, Widen R. Alterations in levels of CD28-/CD8+ suppressor cell precursor and CD45RO+/ CD4+ memory T lymphocytes in the peripheral blood of multiple sclerosis patients. Clin Diagn Lab Immunol 1995; 2:249–252.
8. Najafian N, Chitnis T, Salama AD,  Zhu B, Benou C, Yuan X, et al. Regulatory functions of CD8+CD28- T cells in an autoimmune disease model. J Clin Invest 2003; 112:1037–1048.
9. Ménager-Marcq I, Pomié C, Romagnoli P, Van Meerwijk JP. CD8+CD28- regulatory T lymphocytes prevent experimental inflammatory bowel disease in mice. Gastroenterology 2006; 131:1775–1785.
10. Ben-David H, Sharabi A, Dayan M, Sela M, Mozes E. The role of CD8+CD28-regulatory cells in suppressing myasthenia gravis-associated responses by a dual altered peptide ligand. Proc Natl Acad Sci USA 2007; 104:17459– 17464.
11. Tulunay A, Yavuz S, Direskeneli H, Eksioglu-Demiralp E. CD8+CD28-, suppressive T cells in systemic lupus erythematosus. Lupus 2008; 17:630– 637.
12. North ME, Webster AD, Farrant J. Primary defect in CD8+ lymphocytes in the antibody deficiency disease (common variable immunodeficiency):  abnormalities in intracellular production of interferon-gamma (IFN-c) in CD28+ (‘‘cytotoxic”) and CD28- (‘‘suppressor”) CD8+ subsets. Clin Exp Immunol 1998; 11:70–75.
13. Liu W, Putnam AL, Xu-yu Z,  Szot GL, Lee MR, Zhu S, et al. CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells. J Exp Med 2009; 203:1701–1711.
14. Michalek J, Vrabelova Z, Hrotekova Z, Kyr M, Pejchlova M, Kolouskova S, et al. Immune regulatory T cells in siblings of children suffering from type 1 diabetes mellitus. Scand J Immunol 2006; 64: 531–535.
15. Expanded Disability Status Scale. 2008. Available at: http://www. mult-sclerosis. org/expandeddisabilitystatusscale. html.
16. Feger U, Luther C, Poeschel S, Melms A, Tolosa E, Wiendl H. Increased frequency of CD4+ CD25+ regulatory T cells in the cerebrospinal fluid but not in the blood of multiple sclerosis patients. Clin Exp Immunol 2007; 147:412–418.
17. Haas J,  Hug A, Viehöver A, Fritzsching B, Falk CS, Filser A, et al. Reduced suppressive effect of CD4+CD25high regulatory T cells on the T cell immune response against myelin oligodendrocyte glycoprotein in patients with multiple sclerosis. Eur J Immunol 2005; 35:3343–3352.
18. Viglietta V, Baecher-Allan C, Howard LW, Hafler D. Loss of functional suppression by CD4+CD25+ regulatory T cells in patients with multiple sclerosis. J Exp Med 2004; 199:971–979.
19. Putheti P,  Pettersson A, Soderstrom M, Link H, Huang YM, et al. Circulating CD4+CD25+T regulatory cells are not altered in multiple sclerosis and unaffected by disease-modulating drugs. J Clin Immunol 2004; 24:155–161.
20. Venken K, Hellings N, Broekmans T, Hensen K, Rummens JL, Stinissen P. Natural naive CD4+CD25+CD127low regulatory T cell  (Treg) development and function are disturbed in multiple sclerosis patients: recovery of memory Treg homeostasis during disease progression. J Immunol  2008; 180:6411-6420.
21. Namdar A, Nikbin B, Ghabaee  M, Bayati  A, Izad M. Effect of IFN-ß therapy on the frequency and function of CD4+CD25+ regulatory T cells and Foxp3 gene expression in relapsing–remitting multiple sclerosis (RRMS):A preliminary study. J Neuroimmunol 2010; 218:120–124.
22. Nikoueinejad H, Sharif MR, Amirzargar A, Mirshafiey A, Einollahi B. Regulatory T cells as a therapeutic tool to induce solid-organ transplant 
tolerance: current clinical experiences. Exp Clin Transplant 2013; 11:379-387.
23. Huan J, Culbertson N, Spencer L, Bartholomew R, Burrows GG, Chou YK. Decreased FOXP3 levels in multiple sclerosis patients. J Neurosci Res 2005; 81:45–52.
24. Correale J, Villa A. Role of CD8+CD25+Foxp3+ regulatory T Cells in multiple sclerosis. Ann Neurol 2010; 67:625-638.
25. Mikulkova Z, Praksova P, Stourac P, Bednarik J, Strajtova L, Pacasova R, et al. Numerical defects in CD8+CD28- T-suppressor lymphocyte population in patients with type 1 diabetes mellitus and multiple sclerosis. Cell Immunol  2010; 262:75-79.
26. Frisullo G, Nociti V, Iorio R, Plantone D, Patanella AK, Tonali PA, et al. CD8(+)Foxp3(+) T cells in peripheral blood of relapsing-remitting multiple sclerosis patients. Hum Immunol 2010; 71:437-441.