2013 International Conference & Exhibition on Nutraceuticals and Functional Foods

Formulated extract from multiple citrus peels impairs immune dendritic cells functions

and attenuates allergic contact hypersentivity

Gold lotion (GL), a formulated product made from the peels of citrus fruits, has many pharmacological activities, such as anti-tumor, antioxidant, and anti-inflammatory activities. In this study, we examine the immunomodulatory effect of GL on lipopolysacchride (LPS) stimulated mouse bone marrow-derived DCs maturation and function. Our experimental results show that GL significantly inhibited the pro-inflammatory cytokines and chemokine expression in LPS-induced DCs in a dose dependent manner. In addition, GL treatment led to inhibit the expression of of MHC and costimulatory molecules, influence the ability to take up antigen and attenuated allo-T cell activating ability of LPS stimaultaed DCs. Furthermore, oral administration of GL attenuates the 2,4-Dinitro-1-fluorobenzene induced contact hypersensitivit (CHS) in vivo. Consequently, molecular studies showed that GL decrease LPS-induced MAPK-ERK, p38 phosphrylation and nuclear translocation of NF-κB p65. Taken together, these findings provide new insight into the immunopharmacological role of GL and used to treat DC mediated-chronic inflammatory disorders or autoimmune diseases.

Introduction of Researchers & Universities

Shiming Li^1,1a, Yi-Chin Lin², Chi-Tang Ho¹, Ping-Yi Lin³, Michiko Suzawa⁴, Hsin-Chieh Wang², Ching-Liang Chu⁵, Der-Yuan Chen^2,6, Chi-Chen Lin^*2,6,7,8,

¹Department of Food Science, Rutgers University, New Brunswick, NJ 08901, USA

^1aColleage of Chemistry and Chemical Engineering, Huanggang Normal University, Hubei, 438000, China

²Institute of Biomedical Science, National Chung-Hsing University, Taichung, Taiwan

³Transplant Medicine & Surgery Research Centre, Changhua Christian Hospital, Changhua, Taiwan

⁴Miyauchi Citrus Research Center, Shigoka-Machi, Takasaki, Gunma, 370-0845, Japan

⁵Graduate Institutes of Immunology, College of Medicine, National Taiwan University, Taipei 100, Taiwan

⁶Department of Medical Research and Education, Taichung Veterans General Hospital, Taichung, Taiwan, Republic of China.

⁷Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital, Changhua, Taiwan

⁸Rong Hsing Research Center for Translational Medicine, National Chung Hsing University

*To whom correspondence should be addressed:

Dr. Chi-Chen Lin, Institute of Biomedical Sciences, College of Life Science, National Chung Hsing University, Taichung, Taiwan FAX: +886-4-23592705, E-Mail: lincc@dragon.nchu.edu.tw

References

[1] Hakim IA, Harris RB, Ritenbaugh C. Citrus peel use is associated with reduced risk of squamous cell carcinoma of the skin. Nutrition and cancer 2000;37: 161-168.

[2] Chutia M, Deka Bhuyan P, Pathak MG, Sarma TC, Boruah P. Antifungal activity and chemical composition of Citrus reticulata Blanco essential oil against phytopathogens from North East India. LWT—Food Science and Technology 2009;42:777–780.

[3] Sawamura M, Thi Minh Tu N, Onishi Y, Ogawa E, Choi HS. Characteristic odor components of Citrus reticulata Blanco (ponkan) cold-pressed oil.Bioscience, Biotechnology and Biochemistry 2004;68:1690–1697.

[4] Espina L, Somolinos M, Lorán S, Conchello P, García D, Pagán R. Chemical composition of commercial citrus fruit essential oils and evaluation of their antimicrobial activity acting alone or in combined processes. Food Control 2011;22:896–902.

[5] Ho, SC. and Lin CC. Investigation of heat treating conditions for enhancing the anti-inflammatory activity of citrus fruit (Citrus reticulata) peels. Journal of agricultural and food chemistr. 2008;56:7976-7982.

[6] Epifano F, Genovese S, Curini M. Auraptene: phytochemical and pharmacological properties. Phytochemistry Research Progress 2008;145-62.

[7] Upadhyay RK, Dwivedi P, Ahmad S. Screening of antibacterial activity of six plant essential oils against pathogenic bacterial strains. Asian J Med Sci 2010;2: 152-8.

[8] Yang EJ, Kim SS, Moon JY, Oh T-H, Baik JS, Lee NH, et al. Inhibitory effects of Fortunella japonica var. margarita and Citrus sunki essential oils on nitric oxide production and skin pathogens. Acta microbiologica et immunologica Hungarica. 2010;57:15-27.

[9] Yi E, Yoo ES. A novel bioactive fabric dyed with unripe Citrus grandis Osbeck extract part 1: dyeing properties and antimicrobial activity on cotton knit fabrics. Textile Research Journal 2010;80:2117-23.

[10] Benavente-Garcia O, Castillo J. Update on uses and properties of citrus flavonoids: new findings in anticancer, cardiovascular, and anti-inflammatory activity. Journal of Agricultural and Food Chemistr. 2008;56:6185-205.

[11] Kurowska EM, Manthey JA, Casaschi A, Theriault AG. Modulation of HepG2 cell net apolipoprotein B secretion by the citrus polymethoxyflavone, tangeretin. Lipids 2004;39:143-51.

[12] Manthey JA, Guthrie N, Grohmann K. Biological properties of citrus flavonoids pertaining to cancer and inflammation. Current Medicinal Chemistry 2001;8: 135-53.

[13] Miyazawa M, Okuno Y, Fukuyama M, Nakamura S-i, Kosaka H. Antimutagenic activity of polymethoxyflavonoids from Citrus aurantium. Journal of Agricultural and Food Chemistry 1999:47:5239-44.

[14] Roza JM, Xian-Liu Z, Guthrie N. Effect of citrus flavonoids and tocotrienols on serum cholesterol levels in hypercholesterolemic subjects. Alternative Therapies in Health and Medicine 2007;13:44.

[15] Xiao H, Yang CS, Li S, Jin H, Ho CT, Patel T. Monodemethylated polymethoxyflavones from sweet orange (Citrus sinensis) peel inhibit growth of human lung cancer cells by apoptosis. Molecular nutrition & food research 2009; 53:398-406.

[16] Lai CS, Li S, Liu CB, Miyauchi Y, Suzawa M, Ho CT, Pan MH. Effective suppression of azoxymethane-induced aberrant crypt foci formation in mice with citrus peel flavonoids. Molecular nutrition & food research 2013;57:551-5.

[17] Pan MH, Li S, Lai CS, Miyauchi Y, Suzawa M, Ho CT. Inhibition of citrus flavonoids on 12-O-tetradecanoylphorbol 13-acetate-induced skin inflammation and tumorigenesis in mice. Food Science and Human Wellness 2012;1:65–73.

[18] Lai CS, Li S, Miyauchi Y, Suzawa M, Ho CT, Pan MH. Potent anti-cancer effects of citrus peel flavonoids in human prostate xenograft tumors. Food Funct 2013;4:944-9.

[19] Etoh T, Kim YP, Hayashi M, Suzawa M, Li S, Ho CT, and Komiyama K. Inhibitory effect of a formulated extract from multiple citrus peels on LPS-induced inflammation in RAW 246.7 macrophages. Functional Foods in Health and Disease 2013;3:242-253.

[20] Granucci F, Zanoni I, Ricciardi-Castagnoli P. Central role of dendritic cells in the regulation and deregulation of immune responses. Cell Mol Life Sci 2008;65: 1683-97.

[21] Steinman RM, Hemmi H. Dendritic cells: translating innate to adaptive immunity. Current Topics in Microbiology and Immunology 2006;311:17–58.

[22] Hivroz C, Chemin K, Tourret M, Bohineust A. Crosstalk between T lymphocytes and dendritic cells. Crit Rev Immunol 2012;32:139-155.

[23] Förster R, Braun A, Worbs T. Lymph node homing of T cells and dendritic cells via afferent lymphatics. Trends Immunol 2012;33:271-280.

[24] P. Hubert, N. Jacobs, J. H. Caberg, J. Boniver, and P. Delvenne, “The cross-talk between dendritic and regulatory T cells: good or evil?” Journal of Leukocyte Biology 2007;82:781–794.

[25] Chen DY, Song PS, Hong JS, Chu CL, Pan IH, Chen YM, Lin CH, Lin SH, Lin CC. Dextromethorphan inhibits activations and functions in dendritic cells. Clin Dev Immunol 2013;125643.

[26] Lin CC, Yu YL, Shih CC, Liu KJ, Ou KL, Hong LZ, Chen JD, Chu CL. A novel adjuvant Ling Zhi-8 enhances the efficacy of DNA cancer vaccine by activating dendritic cells. Cancer Immunol Immunother 2011;60:1019-27.

[27] Huang RY, Yu YL, Cheng WC, OuYang CN, Fu E, Chu CL. Immunosuppressive effect of quercetin on dendritic cell activation and function. J Immuno. 2010;184:6815-21.

[28] Kim GY, Han MG, Song YS et al. Proteoglycan isolated from Phellinus linteus induces toll-like receptors 2- and 4- mediated maturation of murine dendritic cells via activation of ERK, p38, and NF-κB. Biol Pharm Buli 2004;27:1656–1662.

[29] Rescigno M, Martino M, Sutherland CL, Gold MR, Ricciardi- Castagnoli P. Dendritic cell survival and maturation are regulated by different signaling pathways. J Exp Med 1998;188:2175–2180.

[30] Yokoi S, Niizeki H, Iida H, Asada H, Miyagawa S. Adjuvant effect of lipopolysaccharide on the induction of contact hypersensitivity to haptens in mice. J Dermatol Sci 2009;53:120-128.

[31] Geldart T, Illidge T. Anti-CD 40 monoclonal antibody. Leuk Lymph 2005;46: 1105–13.

[32] Sharpe AH, Freeman GJ. The B7-CD28 superfamily. Nat Rev Immunol 2002;2: 116–126.

[33] Teague TK, Marrack P, Kappler JW, Vella AT. IL-6 rescues resting mouse T cells from apoptosis. J Immunol 1997;158:5791–5796.

[34] Dienz O, Rincon M. The effects of IL-6 on CD4 T cell responses. Clin Immunol. 2009;130:27-33.

[35] Yoo JK, Cho JH, Lee SW, Sung YC. IL-12 provides proliferation and survival signals to murine CD4+ T cells through phosphatidylinositol 3-kinase/Akt signaling pathway. J Immunol 2002;169:3637-43.

[36] Estaquier, J., T. Idziorek, W. Zou, D. Emilie, C. M. Farber, J. M. Bourez, J. C. Ameisen. T helper type 1/T helper type 2 cytokines and T cell death: preventive effect of interleukin 12 on activation-induced and CD95 (FAS/APO-1)-mediated apoptosis of CD4+ T cells from human immunodeficiency virus-infected persons. J. Exp. Med 1995;182:1759.

[37] Marth, T., M. Zeitz, B. R. Ludviksson, W. Strober, B. L. Kelsall. Extinction of IL-12 signaling promotes Fas-mediated apoptosis of antigen-specific T cells. J. Immunol 1999;162:7233.

[38] Gasparini C, Feldmann M. NF-κB as a target for modulating inflammatory responses. Curr Pharm Des 2012;18:5735-5745.

[39] Rescigno M, Martino M, Sutherland CL, Gold MR, Ricciardi- Castagnoli P. Dendritic cell survival and maturation are regulated by different signaling pathways. J Exp Med 1998;188:2175–2180.

[40] Kim GY, Han MG, Song YS et al. Proteoglycan isolated from Phellinus linteus induces toll-like receptors 2- and 4- mediated maturation of murine dendritic cells via activation of ERK, p38, and NF-κB. Biol Pharm Bull 2004;27:1656–1662.

[41] Wang J, Wang X, Hussain S, et al. Distinct roles of different NF-kappa B subunits in regulating inflammatory and T cell stimulatory gene expression in dendritic cells. J Immunol 2007;178:6777-6788.

[42] Kidd P. Th1/Th2 balance: the hypothesis, its limitations, and implications for health and disease. Altern Med Rev 2003;8:223–46.

[43] Gately MK, Renzetti LM, Magram J, et al. The interleukin-12/interleukin-12-receptor system : role in normal and pathologic immune responses. Annu Rev Immunol 1998;16:495-521.

[44] Wang B, Feliciani C, Freed I, Cai Q, Sauder DN. Insights into molecular mechanisms of contact hypersensitivity gained from gene knockout studies. J Leukoc Biol 2001;70:185–191.

[45] Granucci F, Zanoni I. Role of Toll like receptor-activated dendritic cells in the development of autoimmunity. Front Biosci 2008;13:4817-26.

[46] Barber GN., Innate immune DNA sensing pathways: STING, AIMII and the regulation of interferon production and inflammatory responses. Curr Opin Immunol 2011;23:10-20.