New insight into the immunomodulatory mechanisms of Tretinoin in NMRI mice

Document Type : Original Article


Department of Microbiology, Veterinary Faculty, Urmia University, Urmia, Iran


Objective(s):Recent evidence have proposed that Tretinoin produced in the gut preferentially promote differentiation of FoxP3+Treg cells but inhibits Th17 lymphocytes, and this may be the main immunomdulatory mechanism of Tretinoin  in vivo. This study was done to investigate the effects of Tretinoin in outbred white mice after challenge with sheep red blood cells (SRBC).
Materials and Methods: Twenty male NMRI-mice randomly allocated in two equal groups. Mice were treated with 1×109 SRBCs emulsified in CFA intraperitoneally twice with one weak interval. Animals were bled 5 days after last injection. Moreover, 48 hr before bleeding time, 1×109 SRBCs were injected into the left hind foot pad of mice. Tretinoin (25 mg/kg-every other day) were intraperitoneally injected into the treatment group from the beginning of the study and continued throughout the study. The levels of anti-SRBC antibody and the specific cellular immune responses were measured by microhemagglutination test and footpad thickness, respectively. Moreover, splenocytes were checked for proliferation rate, respiratory burst, cytokine production and FoxP3+Treg cells frequency.
Results: Tretinoin markedly alleviated cellular immunity and concurrently potentiated humoral immunity after mice challenge with SRBCs. Furthermore, aside from reducing NBT reduction and lymphocyte proliferation, Tretinoin markedly suppressed the secretion of interleukin-17 and conversely, increased the production of interleukin-10. However, the level of IFN-γ and the frequency of FoxP3+Treg cells did not alter significantly.
Conclusion: The in vivo immunomudlatoty effects of Tretinoin may be partly due to immune deviation from pro-inflammatory cytokine interleukin-17 to anti-inflammatory cytokine interleukin-10, but not absolutely depend on the expansion of FoxP3+Treg cells.


1. Pino-Lagos K, Benson MJ, Noelle RJ. Retinoic acid in the immune system. Ann N Y Acad Sci 2008; 1143:170-187.
2. Xu J, Drew PD. 9-Cis-retinoic acid suppresses inflammatory responses of microglia and astrocytes. J Neuroimmunol 2006; 171:135-144. 
3. Klemann C, Raveney BJ, Oki S, Yamamura T. Retinoid signals and Th17-mediated pathology. Nihon Rinsho Meneki Gakkai Kaishi 2009; 32:20-28.
4. Klemann C, Raveney BJ, Klemann AK, Ozawa T, von Horsten S, Shudo K, et al. Synthetic retinoid AM80 inhibits Th17 cells and ameliorates experimental autoimmune encephalomyelitis. Am J Pathol 2009; 174:2234-2245. 
5. Manicassamy S, Pulendran B. Retinoic acid-dependent regulation of immune responses by dendritic cells and macrophages. Semin Immunol 2009; 21:22-27.
6. Ross AC. Vitamin A and retinoic acid in T cell-related immunity. Am J Clin Nutr 2012; 96:1166S-1172S. 
7. Massacesi L, Abbamondi AL, Giorgi C, Sarlo F, Lolli F, Amaducci L. Suppression of experimental allergic encephalomyelitis by retinoic acid. J Neurol Sci 1987; 80:55-64.
8. Nozaki Y, Yamagata T, Sugiyama M, Ikoma S, Kinoshita K, Funauchi M. Anti-inflammatory effect of all-trans-retinoic acid in inflammatory arthritis. Clin Immunol 2006; 119:272-279.
9. Osanai M, Nishikiori N, Murata M, Chiba H, Kojima T, Sawada N. Cellular retinoic acid bioavailability determines epithelial integrity: Role of retinoic acid receptor alpha agonists in colitis. Mol Pharmacol 2007; 71:250-258. 
10. Racke MK, Burnett D, Pak SH, Albert PS, Cannella B, Raine CS, et al. Retinoid treatment of experimental allergic encephalomyelitis. IL-4 production correlates with improved disease course. J Immunol 1995; 154:450-458.
11. Zunino SJ, Storms DH, Stephensen CB. Diets rich in polyphenols and vitamin A inhibit the development of type I autoimmune diabetes in nonobese diabetic mice. J Nutr 2007; 137:1216-1221.
12. Dong C. Mouse Th17 cells: current understanding of their generation and regulation. Eur J Immunol 2009; 39:640-644.
13. O'Connor RA, Taams LS, Anderton SM. Translational mini-review series on Th17 cells: CD4 T helper cells: functional plasticity and differential sensitivity to regulatory T cell-mediated regulation. Clin Exp Immunol 2010; 159:137-147. 
14. Sun CM, Hall JA, Blank RB, Bouladoux N, Oukka M, Mora JR, et al. Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid. J Exp Med 2007; 204:1775-1785. 
15. Eller P, Eller K, Wolf AM, Reinstadler SJ, Tagwerker A, Patsch JR, et al. Atorvastatin attenuates murine anti-glomerular basement membrane glomerulonephritis. Kidney Int 2010; 77:428-435.
16. Kinoshita K, Yoo BS, Nozaki Y, Sugiyama M, Ikoma S, Ohno M, et al. Retinoic acid reduces autoimmune renal injury and increases survival in NZB/W F1 mice. J Immunol 2003; 170:5793-5798.
17. Van YH, Lee WH, Ortiz S, Lee MH, Qin HJ, Liu CP. All-trans retinoic acid inhibits type 1 diabetes by T regulatory (Treg)-dependent suppression of interferon-gamma-producing T-cells without affecting Th17 cells. Diabetes 2009; 58:146-155.
18. Youssef S, Stuve O, Patarroyo JC, Ruiz PJ, Radosevich JL, Hur EM, et al. The HMG-CoA reductase inhibitor, atorvastatin, promotes a Th2 bias and reverses paralysis in central nervous system autoimmune disease. Nature 2002; 420:78-84.
19. Hay FC, Westwood OMR. Practical Immunology. 4th ed.  New Jersey: Wiley-Blackwell; 2002.
20. Müller J, Alföldy P, Lemmel E-M. Nitroblue-tetrazolium test for the functional evaluation of phagocytic cells: A critical analysis of the methodology. Agents Actions 1981; 11:384-390.
21. Hamaliaka A, Novikova I. Nitric oxide production disorders in leukocytes of patients with recurrent furunculosis. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2010; 154:163-167.
22. Nabi AH, Islam LN, Rahman MM, Biswas KB. Polymorphonuclear neutrophil dysfunctions in streptozotocin-induced type 1 diabetic rats. J Biochem Mol Biol 2005; 38:661-667.
23. Aranami T, Yamamura T. Th17 Cells and autoimmune encephalomyelitis (EAE/MS). Allergol Int 2008; 57:115-120.
24. Hofstetter H, Gold R, Hartung HP. Th17 Cells in MS and Experimental Autoimmune Encephalomyelitis. Int MS J 2009; 16:12-18.
25. Kobayashi K, Kaneda K, Kasama T. Immunopathogenesis of delayed-type hypersensitivity. Microsc Res Tech 2001; 53:241-245.
26. Kuerten S, Lehmann PV. The immune pathogenesis of experimental autoimmune encephalomyelitis: lessons learned for multiple sclerosis?. J Interferon Cytokine Res 2011; 31:907-916.
27. El-behi M, Rostami A, Ciric B. Current views on the roles of Th1 and Th17 cells in experimental autoimmune encephalomyelitis. J Neuroimmune Pharmacol 2010; 5:189-197.
28. Murdaca G, Colombo BM, Puppo F. The role of Th17 lymphocytes in the autoimmune and chronic inflammatory diseases. Intern Emerg Med 2011; 6:487-495.  
29. Fletcher JM, Lalor SJ, Sweeney CM, Tubridy N, Mills KH. T cells in multiple sclerosis and experimental autoimmune encephalomyelitis. Clin Exp Immunol 2010; 162:1-11.
30. Korn T, Oukka M, Kuchroo V, Bettelli E. Th17 cells: effector T cells with inflammatory properties. Semin Immunol 2007; 19:362-371.
31. Tigno-Aranjuez JT, Lehmann PV, Tary-Lehmann M. Dissociated induction of cytotoxicity and DTH by CFA and CpG. J Immunother 2009; 32:389-398. 
32. Oukka M. Interplay between pathogenic Th17 and regulatory T cells. Ann Rheum Dis 2007; 66: 87-90.
33. Elias KM, Laurence A, Davidson TS, Stephens G, Kanno Y, Shevach EM, et al. Retinoic acid inhibits Th17 polarization and enhances FoxP3 expression through a Stat-3/Stat-5 independent signaling pathway. Blood 2008; 111:1013-1020.
34. Kagami S, Owada T, Kanari H, Saito Y, Suto A, Ikeda K, et al. Protein geranylgeranylation regulates the balance between Th17 cells and Foxp3+ regulatory T cells. Int Immunol 2009; 21:679-689.
35. Mucida D, Park Y, Kim G, Turovskaya O, Scott I, Kronenberg M, et al. Reciprocal TH17 and regulatory T cell differentiation mediated by retinoic acid. Science 2007; 317:256-260. 
36. Korn T, Reddy J, Gao W, Bettelli E, Awasthi A, Petersen TR, et al. Myelin-specific regulatory T cells accumulate in the CNS but fail to control autoimmune inflammation. Nat Med 2007; 13:423-431. 
37. Asadullah K, Sterry W, Volk HD. Interleukin-10 therapy--review of a new approach. Pharmacol Rev 2003; 55:241-269.
38. Saraiva M, O'Garra A. The regulation of IL-10 production by immune cells. Nat Rev Immunol 2010; 10:170-181.