Chitin Micro Particles Regulate Splenocytes Immune Response in Experimental Autoimmune Encephalomyelitis

  • Sanaz Mami Department of Medical Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Farshid Yeganeh Department of Medical Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Elnaz Farahani Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
  • Ali Anissian Department of Veterinary Pathology, Islamic Azad University, Abhar Branch, Abhar, Iran
  • Mustafa Haji Molla Hoseini Department of Medical Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Keywords: Chitin, Cytokine, Experimental autoimmune encephalomyelitis, Immunomodulation, Transcription factor


Contrasting studies are reported on the induction of IL-10 and IFN-γ via chitin microparticles (CMPs) during immune stimulation. Our previous studies have shown marked protection among CMP treated Leishmania-infected mice via regulated IL-10/IFN-γ response, at the present study, once more, examined the inconsistent responses regarding the immunologic response of CMPS. To verify whether CMPs could indeed up-regulate IL-10/IFN-γ axis, isolated spleen cells from the myelin oligodendrocyte glycoprotein (MOG) induced experimental autoimmune encephalomyelitis (EAE) mice were cultured in the presence of MOG peptide and/or CMPs. The effects of CMPs on IL-10, IFN-γ and IL-17 production were evaluated by Enzyme-linked Immunosorbent Assay (ELISA). Moreover, GATA binding protein 3 (Gata3), T-box transcription factor TBX21 (Tbx21), and RAR-related orphan receptor gamma (RORγT) expressions (real-time PCR) were investigated. MOG alone stimulated the production of IFN-γ (p≤0.004) but not, IL-10 (p≤0.140). MOG/chitin stimulation resulted in a significant increase in IFN-γ and IL-10 levels, respectively; (p≤0.004 and p≤0.003) rather than MOG. Additionally, the expression of Tbx21 (p≤0.001), but not Gata3 (p≤0.08), was increased in the MOG/chitin-treated spleen cells. All in all, CMP supports Gata3 independent IL-10 production and promotes Tbx21 dependent IFN-γ induction. These results, alongside our previous data, indicate that CMPs has particular adjuvant effects.

Author Biography

Ali Anissian, Department of Veterinary Pathology, Islamic Azad University, Abhar Branch, Abhar, Iran

Islamic Azad University,Abhar branch,


1. Philibert T, Lee BH, Fabien N. Current Status and New Perspectives on Chitin and Chitosan as Functional Biopolymers. Appl Biochem Biotechnol 2017; 181(4):1314–7.
2. Brodaczewska K, Donskow-ŁYsoniewska K, Doligalska M. Chitin a key factor in immune regulation: Lesson from infection with fungi and chitin bearing parasites. Acta Parasitol 2015; 60(2):337–44.
3. Shen CR, Juang HH, Chen HS,Yang CJ, Wu CJ,Lee MH, et al. The correlation between chitin and acidic mammalian chitinase in animal models of allergic asthma. Int J Mol Sci 2015; 16(11):27371–7.
4. Shibata Y, Foster LA, Bradfield JF, Myrvik QN. Oral administration of chitin down-regulates serum IgE levels and lung eosinophilia in the allergic mouse. J Immunol 2000; 164(3):1314–21.
5. Da Silva CA, Pochard P, Lee CG, Elias JA. Chitin particles are multifaceted immune adjuvants. Am J Respir Crit Care Med 2010; 182(12):1482–91.
6. Da Silva CA, Chalouni C, Williams A,Hartl D, Lee CG, Elias JA, et al. Chitin Is a Size-Dependent Regulator of Macrophage TNF and IL-10 Production. J Immunol 2009; 182(6):3573–82.
7. Nagatani K,Wang S,Llado V, et al. Chitin microparticles for he control of intestinal inflammation.Inflamm Bowel Dis 2012; 18(9):1698-710.
8. Shibata Y, Foster LA, Kurimoto M,Okamura H,Nakamura RM,Kawajiri K, et al. Immunoregulatory roles of IL-10 in innate immunity: IL-10 inhibits macrophage production of IFN-gamma-inducing factors but enhances NK cell production of IFN-gamma. J Immunol 1998; 161(8):4283–8.
9. Kogiso M, Nishiyama A, Shinohara T,Nakamura M,Mizoguchi E,Misawa Y, et al. Chitin particles induce size-dependent but carbohydrate-independent innate eosinophilia. J Leukoc Biol 2011; 90(1):167–76.
10. Koch BE, Stougaard J, Spaink HP. Keeping track of the growing number of biological functions of chitin and its interaction partners in biomedical research. Glycobiology 2015; 25(5):469–82.
11. Lee CG, Da Silva CA, Dela Cruz CS,Ahangari F,Ma B,Kang MJ,He CH, et al. Role of Chitin and Chitinase/Chitinase-Like Proteins in Inflammation, Tissue Remodeling, and Injury. Annu Rev Physiol 2011; 73:479–501.
12. Hoseini MHM, Moradi M, Alimohammadian MH,Shahgoli VK, Darabi H, Rostami A. Immunotherapeutic effects of chitin in comparison with chitosan against Leishmania major infection. Parasitol Int 2016; 65(2):99-104.
13. Ghotloo S, Hoseini MHM, Alimohammadian MH, Khaze V, Memarnejadian A, Rostami A. Immunomodulatory effects of chitin microparticles on Leishmania major-infected BALB/c mice. Parasitol In 2015; 64(2):219-21;
14. Simmons SB, Pierson ER, Lee SY, Goverman JM. Modeling the heterogeneity of multiple sclerosis in animals. Trends Immunol 2013; 34(8):410–22.
15. Dehghani F, Haji Molla Hoseini M, Memarnejadian A,Yeganeh F,Rezaie AM,Khaze V, et al. Immunomodulatory Activities of Chitin Microparticles on Leishmania major-infected Murine Macrophages. Arch Med Res 2011; 42(7):572–6.
16. Hoseini MHM, Moradi M, Alimohammadian MH, Shahgoli VK,Darabi H,Rostami A. Immunotherapeutic effects of chitin in comparison with chitosan against Leishmania major infection. Parasitol Int 2016; 65(2):99–104.
17. Alimohammadi M, Yeganeh F, Haji Molla Hoseini M. Preliminary Study on Gene Expression of Chitinase-Like Cytokines in Human Airway Epithelial Cell Under Chitin and Chitosan Microparticles Treatment. Inflammation 2016; 39(3):1108-15.
18. Solati J, Asiaei M, Hoseini MHM. Using experimental autoimmune encephalomyelitis as a model to study the effect of prenatal stress on fetal programming. Neurol Res 2012; 34(5):478–83.
19. Ayatollahi AM, Haji Molla Hoseini M, Ghanadian SM,Kosari-Nasab M,Mami F,Yazdiniapoure Z. TAMEC: a new analogue of cyclomyrsinol diterpenes decreases anxiety- and depression-like behaviors in a mouse model of multiple sclerosis. Neurol Res 2017; 39(12):1056-65
20. Stromnes IM, Goverman JM Active induction of experimental allergic encephalomyelitis. Nat Protoc 2006; 1(4):1810–9.
21. Gibson-Corley KN, Boyden AW, Leidinger MR, Lambertz AM,Ofori-Amanfo G,Naumann PWet al. A method for histopathological study of the multifocal nature of spinal cord lesions in murine experimental autoimmune encephalomyelitis. PeerJ 2016; 4:e1600.
22. Lee J,Choi J,Lee W,Ko K,Kim S. .Dehydrodiconiferyl alcohol (DHCA) modulates the differentiation of Th17 and Th1 cells and suppresses experimental autoimmune encephalomyelitis. Mol Immunol 2015; 68(2 Pt B):434–44.
23. Cai Y, Shen H, Qin C,Zhou J, Lai W, Pan J, et al. The Spatio-Temporal Expression Profiles of CD4 + T Cell Differentiation and Function-Related Genes During EAE Pathogenesis. Inflammation 2017; 40(1):195–204.
24. Mandolesi G, Musella A, Gentile A, Grasselli G,Haji N,Sepman H, et al. Interleukin-1 Alters Glutamate Transmission at Purkinje Cell Synapses in a Mouse Model of Multiple Sclerosis. J Neurosci 33(29):12105-21.
25. Zhang H, Qi Y, Yuan Y,Cai L, Xu H, Zhang L, et al. Paeoniflorin Ameliorates Experimental Autoimmune Encephalomyelitis via Inhibition of Dendritic Cell Function and Th17 Cell Differentiation. Sci Rep 2017; 7:41887.
26. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002; 30(9):e36.
27. Frischer JM, Bramow S, Dal-Bianco A, Claudia F. Lucchinett CF,Rauschka H,et al.The relation between inflammation and neurodegeneration in multiple sclerosis brains Brain. 2009; 132(5): 1175–89.
28. Kubo M, Motomura Y. Transcriptional regulation of the anti-inflammatory cytokine IL-10 in acquired immune cells. Front Immuno 2012; 3:275.
29. MacMicking J, Xie Q, Nathan C. Nitric Oxide and Macrophage Function. Annu Rev Immunol 1997; 15:323–350.
30. Tripathi P, Tripathi P, Kashyap L, Singh V. The role of nitric oxide in inflammatory reactions. FEMS Immunol Med Microbiol 2007; 51(3):443–52.
31. ’t Hart BA, Gran B, Weissert R. EAE: Imperfect but useful models of multiple sclerosis. Trends Mol Med 2011; 17(3):119–25.
32. Becker K, Aimanianda V, Wang X, Gresnigt MS, Ammerdorffer A, Jacobs CW, et al. Aspergillus Cell Wall Chitin Induces Anti- and Proinflammatory cytokines in human PBMCs via the Fc-gamm receptor/Syk/PI3K pathway. MBio (2016); 7(3):1–11.
33. Bueter CL, Lee CK, Rathinam VAK,Healy GJ, Taron CH, Specht CA, et al. Chitosan but not chitin activates the inflammasome by a mechanism dependent upon phagocytosis. J Biol Chem 2011; 286(41):35447-55.
34. Diamantstein T, Klos M, Osawa H, Chen ZC. Chitin: an immunological adjuvant and a polyclonal B-lymphocyte activator. Int Arch Allergy Appl Immunol 1982; 68(4):377–81.
35. Matsushita T, Yanaba K, Bouaziz J-D, Fujimoto M, Tedder TF. Regulatory B cells inhibit EAE initiation in mice while other B cells promote disease progression. J Clin Invest 2008; 118(10):3420–30.
36. Reese TA, Liang H, Tager AM,Andrew D. Luster, Nico Van Rooijen, David Voehringer, et al. Chitin Induces Tissue Accumulation of Innate Immune Cells Associated with Allergy Tiffany. Nature 2007; 447(7140):92–6.
37. Dubey LK, Moeller JB, Schlosser A,Sorensen GL, Holmskov U. Chitin enhances serum IgE in Aspergillus fumigatus induced allergy in mice. Immunobiology 2015; 220(6):714–21.
38. Wagener J, Malireddi RKS, Lenardon MD,Köberle M,Vautier S,MacCallum DM, et al Fungal Chitin Dampens Inflammation through IL-10 Induction Mediated by NOD2 and TLR9 Activation. PLoS Pathog 2014; 10(4):e1004050.
39. Murray PJ, Wang L, Onufryk C, Tepper RI, Young RA. T cell-derived IL-10 antagonizes macrophage function in mycobacterial infection. J Immunol 1997; 158(1):315–21.
40.Rutz S,.Janke M,.Kassner N,Hohnstein.T,.Krueger M,.Scheffold A. Notch regulates IL-10 production by T helper 1 cells.Proc Natl Acad Sci U S A 2008; 105(9):3497-502.
41. Kahn D a, Archer DC, Gold DP, Kelly CJ. .Adjuvant immunotherapy is dependent on inducible nitric oxide synthase. J Exp Med 2001; 193:1261–8.
42. Lubina-Dąbrowska N, Stepień A, Sulkowski G, Dąbrowska-Bouta B, Langfort J, Chalimoniuk M. Effects of IFN-β1a and IFN-β1b treatment on the expression of cytokines, inducible NOS (NOS type II), and myelin proteins in animal model of multiple sclerosis. Arch Immunol Ther Exp (Warsz) 2017; 65(4):325–38.
43. Gold DP, Schroder K, Powell HC, Kelly CJ. Nitric oxide and the immunomodulation of experimental allergic encephalomyelitis. Eur J Immunol 1997; 27(11):2863–9.
44. O’Connor RA, Li X, Blumerman S, Anderton SM, Noelle RJDalton DK. Adjuvant Immunotherapy of Experimental Autoimmune Encephalomyelitis: Immature Myeloid Cells Expressing CXCL10 and CXCL16 Attract CXCR3+CXCR6+ and Myelin-Specific T Cells to the Draining Lymph Nodes Rather Than the Central Nervous System. J Immunol 2012; 188(5):2093–2101.
How to Cite
Mami S, Yeganeh F, Farahani E, Anissian A, Haji Molla Hoseini M. Chitin Micro Particles Regulate Splenocytes Immune Response in Experimental Autoimmune Encephalomyelitis. Iran J Allergy Asthma Immunol. 18(2):190-199.
Original Article(s)