Vitamin C inhibited FasL-induced apoptotic death of mouse dendritic cells through c-FLIP expression

Nguyen Thi Xuan; Le Thi Thu Hien

doi:10.15625/1811-4989/16/4/13108

Author affiliations

Authors

Nguyen Thi Xuan Institute of Genome Research, Vietnam Academy of Science and Technology http://orcid.org/0000-0003-3494-5136
Le Thi Thu Hien Institute of Genome Research, Vietnam Academy of Science and Technology

DOI:

https://doi.org/10.15625/1811-4989/16/4/13108

Keywords:

c-FLIP, dendritic cell, Fas ligand, phosphatidylserine, vitamin C

Abstract

Vitamin C (VitC) is a potent antioxidant and contributes as an apoptosis inhibitor by preventing death receptor-triggered caspase 8 activity. Fas ligand (FasL) induces the apoptotic cell death via activation of Fas signaling, which is dependent on the expression level of anti-apoptotic molecule c-FLIP (FADD-like IL-1beta-converting enzyme-inhibitory proteins). The present study addressed the effects of VitC on survival of dendritic cells (DCs), a regulator of innate and adaptive immunity. To this end, mouse bone marrow cells were isolated and cultured to attain bone marrow-derived DCs (BMDCs). The cells were treated with FasL in the presence or absence of VitC. Real time RT-PCR, Western blotting and FACS analysis were performed to determine different hallmarks of DC apoptosis. As a result, FasL treatment resulted in activation of caspase 8 and stimulation of cell membrane scrambling, the effects were supressed when VitC was present in the cell culture or the cells were transfected with FLIP siRNA. In conclusion, VitC prevented FasL-triggered DC apoptosis mediated through the expression of c-FLIP.

Downloads

Download data is not yet available.

References

Ashany D, Savir A, Bhardwaj N, Elkon KB (1999) Dendritic cells are resistant to apoptosis through the Fas (CD95/APO-1) pathway. J Immunol 163: 5303–11.

Ashany D, Song X, Lacy E, Nikolic-Zugic J, Friedman SM, Elkon KB (1995) Th1 CD4+ lymphocytes delete activated macrophages through the Fas/APO-1 antigen pathway. Proc Natl Acad Sci USA 92: 11225–9.

Buonocore S, Van Meirvenne S, Demoor FX, Paulart F, Thielemans K, Goldman M, et al (2002) Dendritic cells transduced with viral interleukin 10 or Fas ligand: no evidence for induction of allotolerance in vivo. Transplantation 73: S27–30.

Conti S, Petrungaro S, Marini ES, Masciarelli S, Tomaipitinca L, Filippini A, et al (2016) A novel role of c-FLIP protein in regulation of ER stress response. Cell Signal 28: 1262–1269.

Esmailzadeh S, Huang Y, Su MW, Zhou Y, Jiang X (2015) BIN1 tumor suppressor regulates Fas/Fas ligand-mediated apoptosis through c-FLIP in cutaneous T-cell lymphoma. Leukemia 29: 1402–13.

Golan-Gerstl R, Wallach-Dayan SB, Zisman P, Cardoso WV, Goldstein RH, Breuer R (2012) Cellular FLICE-like inhibitory protein deviates myofibroblast fas-induced apoptosis toward proliferation during lung fibrosis. Am J Respir Cell Mol Biol 47: 271–9.

Hamai A, Richon C, Meslin F, Faure F, Kauffmann A, Lecluse Y, et al (2006) Imatinib enhances human melanoma cell susceptibility to TRAIL-induced cell death: Relationship to Bcl-2 family and caspase activation. Oncogene 25: 7618–34.

Haverkamp JM, Smith AM, Weinlich R, Dillon CP, Qualls JE, Neale G, et al (2014) Myeloid-derived suppressor activity is mediated by monocytic lineages maintained by continuous inhibition of extrinsic and intrinsic death pathways. Immunity 41: 947–59.

Jaita G, Zarate S, Ferraris J, Gottardo MF, Eijo G, Magri ML, et al (2016) Estradiol upregulates c-FLIPlong expression in anterior pituitary cells. Horm Metab Res 48: 275–9.

Jeong CH, Joo SH 2016 Downregulation of reactive oxygen species in apoptosis. J Cancer Prev 21: 13–20.

Jeong YJ, Kim JH, Hong JM, Kang JS, Kim HR, Lee WJ, et al (2014) Vitamin C treatment of mouse bone marrow-derived dendritic cells enhanced CD8(+) memory T cell production capacity of these cells in vivo. Immunobiology 219: 554–64.

Kim HW, Cho SI, Bae S, Kim H, Kim Y, Hwang YI, et al (2012) Vitamin C up-regulates expression of CD80, CD86 and MHC class II on dendritic cell line, DC-1 via the activation of p38 MAPK. Immune Netw 12: 277–83.

Levy R, Shriker O, Porath A, Riesenberg K, Schlaeffer F (1996) Vitamin C for the treatment of recurrent furunculosis in patients with imparied neutrophil functions. J Infect Dis 173: 1502–5.

Liu KJ, Lee YL, Yang YY, Shih NY, Ho CC, Wu YC, et al (2011) Modulation of the development of human monocyte-derived dendritic cells by lithium chloride. J Cell Physiol 226: 424–33.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–8.

Llanos C, Carreno LJ, Kalergis AM (2011) Contribution of dendritic cell/T cell interactions to triggering and maintaining autoimmunity. Biol Res 44: 53–61.

Lucken-Ardjomande S, Martinou JC (2005) Regulation of Bcl-2 proteins and of the permeability of the outer mitochondrial membrane. C R Biol 328: 616–31.

Martino A (2007) Sphingosine 1-phosphate as a novel immune regulator of dendritic cells. J Biosci 32: 1207–12.

Micheau O, Lens S, Gaide O, Alevizopoulos K, Tschopp J (2001) NF-kappaB signals induce the expression of c-FLIP. Mol Cell Biol 21: 5299–305.

Ouaaz F, Arron J, Zheng Y, Choi Y, Beg AA (2002) Dendritic cell development and survival require distinct NF-kappaB subunits. Immunity 16: 257–70.

Perez-Cruz I, Carcamo JM, Golde DW (2003) Vitamin C inhibits FAS-induced apoptosis in monocytes and U937 cells. Blood 102: 336–43.

Richardson BC, Lalwani ND, Johnson KJ, Marks RM (1994) Fas ligation triggers apoptosis in macrophages but not endothelial cells. Eur J Immunol 24: 2640–5.

Shumilina E, Zahir N, Xuan NT, Lang F (2007) Phosphoinositide 3-kinase dependent regulation of Kv channels in dendritic cells. Cell Physiol Biochem 20: 801–8.

Sun J, Luo H, Nie W, Xu X, Miao X, Huang F, et al

(2015) Protective effect of RIP and c-FLIP in preventing liver cancer cell apoptosis induced by TRAIL. Int J Clin Exp Pathol 8: 6519–25.

Suss G, Shortman K (1996) A subclass of dendritic cells kills CD4 T cells via Fas/Fas-ligand-induced apoptosis. J Exp Med 183: 1789–96.

Tschopp J, Irmler M, Thome M (1998) Inhibition of fas death signals by FLIPs. Curr Opin Immunol 10: 552–8.

Vissers MC, Hampton MB (2004) The role of oxidants and vitamin C on neutrophil apoptosis and clearance. Biochem Soc Trans 32: 499–501.

Wajant H (2003) Targeting the FLICE Inhibitory Protein (FLIP) in cancer therapy. Mol Interv 3: 124–7.

Wu YJ, Wu YH, Mo ST, Hsiao HW, He YW, Lai MZ (2015) Cellular FLIP inhibits myeloid cell activation by suppressing selective innate signaling. J Immunol 195: 2612–23.

Xuan NT, Shumilina E, Schmid E, Bhavsar SK, Rexhepaj R, Gotz F, et al (2010) Role of acidic sphingomyelinase in thymol-mediated dendritic cell death. Mol Nutr Food Res 54: 1833–41.