Evaluation of the Effect of the 47 kDa Protein Isolated from Aged Garlic Extract on Dendritic Cells

Document Type: Original Article

Authors

Department of Immunology, School of Medicine, Tarbiat Modares University, Tehran, Iran

Abstract

Objective(s)
 Garlic (Allium sativum) is known as a potent spice and a medicine with broad therapeutic properties ranging from antibacterial to anticancer, and anticoagulant. One of the major purified garlic protein components is the 47 kDa protein. In this study, the effect of 47 kDa protein extracted from aged garlic (AGE) was evaluated on mouse dendritic cell (DC) maturation in vitro.
Materials and Methods
Forty seven kDa protein was purified from AGE by ammonium sulfate precipitation and gel filtration. SDS-PAGE was used to determine the molecular weight and purity of the isolated protein. DCs were purified from spleen of BALB/c mice by Nycodenz centrifugation and their adhesiveness to the plastic dish. The 47 kDa protein isolated from AGE was added to DCs medium during the overnight culture and the expression of DC surface markers was assessed via flowcytometry.
Results
The 47 kDa protein-treated DCs lowered the expression of DC maturation markers including: CD40, CD86 and MHC-II in comparison with non-treated DCs; (median of 41% versus 47%, 84% versus 91% and 83% versus 90%, respectively) but we observed no statistical difference between the two groups.
Conclusion
Upon treatment with DCs with 47 kDa protein, DCs down regulated the expression of costimulatory and MHC-II surface molecules, which is similar to tolerogenic DC phenotype. According to the results of the present study, we found that 47 kDa protein purified from AGE can be considered as a potential candidate to generate tolerogenic DCs in vitro.
 

Keywords


1. Iciek M, Kwiecien I, Wlodek L. Biological properties of garlic and garlic-derived oraganosulfur compound. Environ Mol Mutagen 2009; 50:247-265.

2. Morihara N, Ide N, Weiss N. Aged garlic extract inhibits CD36 expression in human macrophages via modulation of the PPARγ pathway. Phytother Res 2010; 24:602-608.

3. Campbell JH, Efendy JL, Smith NJ, Campbell GR. Molecular basis by which garlic suppresses atherosclerosis. J Nutr 2001;131:1006-1009.

4. Lau BH, Yamasaki T, Gridley DS. Garlic compounds modulate macrophage and T-lymphocyte functions. Mol Biother 1991; 3:103-107.

5. Sigounas G, Hooker J, Anagnostou A, Steiner M. S-allylmercaptocysteine inhibits cell proliferation and reduces the viability of erythroleukemia, breast, and prostate cancer cell lines. Nutr Cancer 1997; 27:186-191.

6. Singh SV, Powolny AA, Stan SD, Xiao D, Arlotti JA, Warin R, et al. Garlic constituent diallyl trisulfide prevents development of poorly differentiated prostate cancer and pulmonary metastasis multiplicity in TRAMP mice. Cancer Res 2008;  68:9503-9511.

7. Katsuki T, Hirata K, Ishikawa H, Matsuura N, Sumi S, Itoh H. Aged garlic extract has chemopreventative effects on 1,2-dimethylhydrazine-induced colon tumors in rats. J Nutr 2006; 136:847-851.

8. Hassan ZM, Yaraee R, Zare N, Ghazanfari T, Sarraf Nejad AH, Nazori B. Immunomodulatory affect of R10 fraction of garlic extract on natural killer activity. Int Immunopharmacol 2003; 3:1483-1489.

9. Ghazanfari T, Hassan ZM, Ebrahimi M. Immunomodulatory activity of a protein isolated from garlic extract on delayed type hypersensitivity. Int Immunopharmacol 2002; 2:1541-1549.

10.Ghazanfari T, Hassan ZM, Ebtekar M, Ahmadiani A, Naderi G, Azar A. Garlic induces a shift in cytokine pattern in Leishmania major-infected BALB/c mice. Scand J Immunol  2000; 52:491-495.

11. Chandrashekar PM, Venkatesh YP. Identification of the protein components displaying immunomodulatory activity in aged garlic extract. J Ethnopharmacol  2009; 124:384-390.

12. Kasuga S, Uda N, Kyo E, Ushijima M, Morihara N, Itakura Y. Pharmacologic activities of aged garlic extract in comparison with other garlic preparations. J Nutr 2001; 131:1080-1084.

13. Zare A, Farzaneh P, Pourpak Z, Zahedi F, Moin M, Shahabi S, et al. Purified aged garlic extract modulates allergic airway inflammation in BALB/c mice. Iran J Allergy Asthma Immunol 2008; 7:133-141.

14. Kyo E, Uda N, Kasuga S, Itakura Y. Immunomodulatory effects of aged garlic extract. J Nutr 2001; 131:1075-1079.

15. Daneshmandi S, Hajimoradi M, Ahmadabad HN, Hassan ZM, Roudbary M, Ghazanfari T. Effect of 14-kDa and 47-kDa protein molecules of age garlic extract on peritoneal macrophages. Immunopharmacol Immunotoxicol 2011;  33:21-27.

16. Harry RA, Anderson AE, Isaacs JD, Hilkens CM. Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis. Ann Rheum Dis 2010; 69:2042-2050.

17. Valentinis B, Bianchi A, Zhou D, Cipponi A, Catalanotti F, Russo V, et al. Direct effects of polymyxin B on human dendritic cells maturation. The role of IkappaB-alpha/NF-kappaB and ERK1/2 pathways and adhesion. J Biol Chem 2005;   280:14264-14271.

18. Vremec D, Zorbas M, Scollay R, Saunders DJ, Ardavin CF, Wu L, et al. The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells. J Exp Med 1992; 176:47-58.

19. Zheng X, Suzuki M, Ichim TE, Zhang X, Sun H, Zhu F, et al. Treatment of autoimmune arthritis using RNA interference-modulated dendritic cells. J Immunol 2010; 184:6457-6464.

20. Thomson AW, Robbins PD. Tolerogenic dendritic cells for autoimmune disease and transplantation. Ann Rheum Dis  2008; 67:90-96.

21. Anderson AE, Sayers BL, Haniffa MA, Swan DJ, Diboll J, Wang XN, et al. Differential regulation of naive and memory CD4+ T cells by alternatively activated dendritic cells. J Leukoc Biol  2008; 84:124-133.

22. Salazar L, Aravena O, Abello P, Escobar A, Contreras-Levicoy J, Rojas-Colonelli N, et al. Modulation of established murine collagen-induced arthritis by a single inoculation of short-term lipopolysaccharide-stimulated dendritic cells. Ann Rheum Dis  2008; 67:1235-1241.

23. Popov I, Li M, Zheng X, San H, Zhang X, Ichim TE, et al. Preventing autoimmune arthritis using antigen-specific immature dendritic cells: a novel tolerogenic vaccine. Arthritis Res Ther  2006;8:141-151

24.  Charbonnier LM, van Duivenvoorde LM, Apparailly F, Cantos C, Han WG, Noel D, et al. Immature dendritic cells suppress collagen-induced arthritis by in vivo expansion of CD49b+ regulatory T cells. J Immunol 2006; 177:3806-3813.

25.  Pedersen AE, Gad M, Walter MR, Claesson MH. Induction of regulatory dendritic cells by dexamethasone and 1alpha,25-Dihydroxyvitamin D(3). Immunol Let t 2004; 91:63-69.

26.  Ludewig B, Ochsenbein AF, Odermatt B, Paulin D, Hengartner H, Zinkernagel RM. Immunotherapy with dendritic cells directed against tumor antigens shared with normal host cells results in severe autoimmune disease. J Exp Med 2000; 191:795-804.