Hepatic stellate cells promote immunotolerance following orthotopic liver transplantation in rats via induction of T cell apoptosis and regulation of Th2/Th3‑like cell cytokine production

Jiang,Zhijun; Chen,Ying; Feng,Xiaonin; Jiang,Jianwen; Chen,Tianxiang; Xie,Haiyang; Zhou,Lin; Zheng,Shusen

doi:10.3892/etm.2012.801

Hepatic stellate cells promote immunotolerance following orthotopic liver transplantation in rats via induction of T cell apoptosis and regulation of Th2/Th3‑like cell cytokine production

Authors:
- Zhijun Jiang
- Ying Chen
- Xiaonin Feng
- Jianwen Jiang
- Tianxiang Chen
- Haiyang Xie
- Lin Zhou
- Shusen Zheng
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Affiliations: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, P.R. China
Published online on: November 6, 2012 https://doi.org/10.3892/etm.2012.801
Pages: 165-169

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Abstract

Hepatic stellate cells (HSCs) have been demonstrated to have immunoinhibitory activity. The aim of this study was to investigate the role of HSCs in the development of immunotolerance following liver transplantation. A rat liver transplantation tolerance model [donor Lewis into recipient Dark Agouti (DA)] and rejection (donor DA into recipient Lewis) was established. On the 7th day following transplantation, the HSCs and T cells were isolated from the rats of either the tolerance or rejection group and cultured together. The apoptosis rate of the T cells was determined 24 h later by flow cytometry following staining with anti‑CD3 mAb and Annexin V‑FITC/PI. Additionally, the FasL expression of HSCs was determined by flow cytometry following staining with anti‑FasL mAb. The protein levels of IL‑2, TNF‑α, TGF‑β and IL‑10 in the supernatant collected from mixed lymphocyte reaction cultures of HSCs and T cells for 5 days were measured using ELISA assays. HSCs isolated from the tolerance group had a higher T‑cell apoptosis induction activity compared with those of the rejection group. The activity of the HSCs was partially reversed by FasL blocking mAb. Accordingly, the FasL expression level of HSCs in the tolerance group was revealed to be higher than that of the rejection group. Moreover, HSCs stimulated IL‑10 and TGF‑β production in the tolerance group. This study suggests that HSCs are involved in liver transplantation immune tolerance via the induction of T‑cell apoptosis partially mediated by the Fas/FasL pathway and the activation of Th2/Th3‑like cell cytokine production.

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1	Orlando G, Soker S and Wood K: Operational tolerance after liver transplantation. J Hepatol. 50:1247–1257. 2009. View Article : Google Scholar
2	Zhong R, He G, Sakai Y, Li XC, Garcia B, Wall W, Duff J, Stiller C and Grant D: Combined small bowel and liver transplantation in the rat: possible role of the liver in preventing intestinal allograft rejection. Transplantation. 52:550–552. 1991. View Article : Google Scholar : PubMed/NCBI
3	Wang C, Sun J, Wang L, Li L, Horvat M and Sheil R: Combined liver and pancreas transplantation induces pancreas allograft tolerance. Transplant Proc. 29:1145–1146. 1997. View Article : Google Scholar : PubMed/NCBI
4	Sarnacki S, Révillon Y, Cerf-Bensussan N, Calise D, Goulet O and Brousse N: Long-term small-bowel graft survival induced by a spontaneously tolerated liver allograft in inbred rat strains. Transplantation. 54:383–385. 1992. View Article : Google Scholar : PubMed/NCBI
5	Bumgardner GL and Orosz CG: Unusual patterns of alloimmunity evoked by allogeneic liver parenchymal cells. Immunol Rev. 174:260–279. 2000. View Article : Google Scholar : PubMed/NCBI
6	Bumgardner GL, Heininger M, Li J, Xia D, Parker-Thornburg J, Ferguson RM and Orosz CG: A functional model of hepatocyte transplantation for in vivo immunologic studies. Transplantation. 65:53–61. 1998. View Article : Google Scholar : PubMed/NCBI
7	Tiegs G and Lohse AW: Immune tolerance: what is unique about the liver. J Autoimmun. 34:1–6. 2010. View Article : Google Scholar : PubMed/NCBI
8	Winau F, Hegasy G, Weiskirchen R, Weber S, Cassan C, Sieling PA, Modlin RL, Liblau RS, Gressner AM and Kaufmann SH: Ito cells are liver-resident antigen-presenting cells for activating T cell responses. Immunity. 26:117–129. 2007. View Article : Google Scholar : PubMed/NCBI
9	Yu MC, Chen CH, Liang X, Wang L, Gandhi CR, Fung JJ, Lu L and Qian S: Inhibition of T cell responses by hepatic stellate cells via B7-H1-mediated T-cell apoptosis in mice. Hepatology. 40:1312–1321. 2004. View Article : Google Scholar : PubMed/NCBI
10	Chen CH, Kuo LM, Chang Y, Wu W, Goldbach C, Ross MA, Stolz DB, Chen L, Fung JJ, Lu L and Qian S: In vivo immune modulatory activity of hepatic stellate cells in mice. Hepatology. 44:1171–1181. 2006. View Article : Google Scholar : PubMed/NCBI
11	Chen CH, Shu KH, Su YH, Tang KY, Cheng CH, Wu MJ, Yu TM, Chuang YW and Hu C: Cotransplantation of hepatic stellate cells attenuates the severity of graft-versus-host disease. Transplant Proc. 42:971–975. 2010. View Article : Google Scholar : PubMed/NCBI
12	Kamada N: The immunology of experimental liver transplantation in the rat. Immunology. 55:369–389. 1985.PubMed/NCBI
13	Kamada N and Calne RY: Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage. Transplantation. 28:47–50. 1979. View Article : Google Scholar : PubMed/NCBI
14	Niki T, Pekny M, Hellemans K, Bleser PD, Berg KV, Vaeyens F, Quartier E, Schuit F and Geerts A: Class VI intermediate filament protein nestin is induced during activation of rat hepatic stellate cells. Hepatology. 29:520–527. 1999. View Article : Google Scholar : PubMed/NCBI
15	Iredale JP, Benyon RC, Pickering J, McCullen M, Northrop M, Pawley S, Hovell C and Arthur MJ: Mechanisms of spontaneous resolution of rat liver fibrosis hepatic stellate cell apoptosis and reduced hepatic expression of metalloproteinase inhibitors. J Clin Invest. 102:538–549. 1998. View Article : Google Scholar
16	Lang A, Schoonhoven R, Tuvia S, Brenner DA and Rippe RA: Nuclear factor kappaB in proliferation, activation, and apoptosis in rat hepatic stellate cells. J Hepatol. 33:49–58. 2000. View Article : Google Scholar : PubMed/NCBI
17	Liu C, Gaça MD, Swenson ES, Vellucci VF, Reiss M and Wells RG: Smads 2 and 3 are differentially activated by transforming growth factor-beta (TGF-beta) in quiescent and activated hepatic stellate cells. Constitutive nuclear localization of Smads in activated cells is TGF-beta-independent. J Biol Chem. 278:11721–11728. 2003. View Article : Google Scholar
18	Qian S, Lu L, Fu F, Li Y, Li W, Starzl TE, Fung JJ and Thomson AW: Apoptosis within spontaneonsly accepted mouse liver allografts: evidence for deletion of cytotoxic T cells and implications for tolerance induction. J Immunol. 158:4654–4661. 1997.
19	Griffith TS, Brunner T, Fletcher SM, Green DR and Ferguson TA: Fas ligand-induced apoptosis as a mechanism of immune privilege. Science. 270:1189–1192. 1995. View Article : Google Scholar : PubMed/NCBI
20	Garnier H, Clot JP, Bertrand M, Camplez P, Kunlin A, Gorin JP, Le Goaziou F, Lévy R and Cordier G: Liver transplantation in the pig: surgical approach. C R Acad Sci Hebd Seances Acad Sci D. 260:5621–5623. 1965.(In French).
21	Kamada N, Brons G and Davies HS: Fully allogeneic liver grafting in rats induces a state of systemic nonreactivity to donor transplantation antigens. Transplantation. 29:429–431. 1980. View Article : Google Scholar
22	Qian S, Demetris AJ, Murase N, Rao AS, Fung JJ and Starzl TE: Murine liver allograft transplantation: tolerance and donor cell chimerism. Hepatology. 19:916–924. 1994. View Article : Google Scholar : PubMed/NCBI
23	Fairchild PJ and Waldmann H: Dendritic cells and prospects for transplantation tolerance. Curr Opin Immunol. 12:528–535. 2000. View Article : Google Scholar : PubMed/NCBI
24	Sun Z, Wada T, Maemura K, Uchikura K, Hoshino S, Diehl AM and Klein AS: Hepatic allograft-derived Kupfer cells regulate T cell response in rats. Liver Transpl. 9:489–497. 2003. View Article : Google Scholar : PubMed/NCBI
25	Limmer A and Knolle PA: Liver sinusoidal endothelial cells: a new type of organ-resident antigen-presenting cell. Arch Immunol Ther Exp (Warsz). 49(Suppl 1): S7–S11. 2001.PubMed/NCBI
26	Pan TL, Goto S, Lin YC, Lord R, Chiang KC, Lai CY, Chen YS, Eng HL, Cheng YF, Tatsuma T, Kitano S, Lin CL and Chen CL: The fas and fas 1igand pathways in liver allograft tolerance. Clin Exp Immunol. 118:180–187. 1999. View Article : Google Scholar : PubMed/NCBI
27	Rivero M, Crespo J, Mayorga M, et al: Involvement of the Fas system in liver allograft rejection. Am J Gastroenterol. 97:1501–1506. 2002. View Article : Google Scholar : PubMed/NCBI
28	Selenko-Gebauer N, Majdic O, Szekeres A, Höfler G, Guthann E, Korthäuer U, Zlabinger G, Steinberger P, Pickl WF, Stockinger H, Knapp W and Stöckl J: B7-H1 (Programmed death-1 ligand) on dendritic cells is involved in the induction and maintenance of T cell anergy. J Immunol. 170:3637–3644. 2003. View Article : Google Scholar : PubMed/NCBI
29	Dong H, Strome SE, Salomao DR, Tamura H, Hirano F, Flies DB, Roche PC, Lu J, Zhu G, Tamada K, Lennon VA, Celis E and Chen L: Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med. 8:793–800. 2002. View Article : Google Scholar : PubMed/NCBI
30	Lu L, Qian S, Hershberger PA, Rudert WA, Lynch DH and Thomson AW: Fas ligand (CD95L) and B7 expression on dendritic cells provide counterregulatory signals for T cell survival and proliferation. J Immunol. 158:5676–5684. 1997.PubMed/NCBI
31	Savage CO, Hughes CC, Pepinsky RB, Wallner BP, Freedman AS and Pober JS: Endothelial cell lymphocyte function-associated antigen-3 and an unidentified ligand act in concert to provide costimulation to human peripheral blood CD4+ T cells. Cell Immunol. 137:150–163. 1991.PubMed/NCBI
32	Chen L: B7-H1 connection of innate and adoptive immunity against tumor dormancy. Blood. 105:2242–2243. 2005. View Article : Google Scholar
33	Subudhi SK, Alegre ML and Fu YX: The balance of immune responses: costimulation verse coinhibition. J Mol Med (Berl). 83:193–202. 2005. View Article : Google Scholar : PubMed/NCBI
34	Fernandes H, Koneru B, Fernandes N, Hameed M, Cohen MC, Raveche E and Cohen S: Investigation of promoter polymorphisms in the tumor necrosis factor-alpha and interleukin-10 gene in liver transplant patients. Transplantation. 73:1886–1891. 2002. View Article : Google Scholar : PubMed/NCBI
35	Karrar A, Broomé U, Uzunel M, Qureshi AR and Sumitran-Holgersson S: Human liver sinusoidal endothelial cells induce apoptosis in activated T cells: a role in tolerance induction. Gut. 56:243–252. 2007. View Article : Google Scholar : PubMed/NCBI