Effects of human umbilical cord mesenchymal stem cell therapy on the immune function and prognosis in patients with decompensated liver cirrhosis due to hepatitis B

doi:10.3969/j.issn.2095-4344.2017.17.012

Abstract

Abstract:

BACKGROUND: A large number of experiments in vivo and in vitro have shown that mesenchymal stem cells may obviously inhibit the lymphocytes and other immunocytes.

OBJECTIVE: To investigate the effect of human umbilical cord mesenchymal stem cell transplantation on the immune function and prognosis of patients suffering decompensated liver cirrhosisn due to hepatitis B.

METHODS: 118 patients with decompensated cirrhosis due to hepatitis B were randomly divided into control group (n=59) and observation group (n=59). The two groups all received normal medical treatment, and in addition, the observation group also received human umbilical cord mesenchymal stem cell transplantation. (4.0-4.5)×10⁸ stem cells were transplanted twice by intervention via proper hepatic artery (10 mL) and intravenous infusion (10 mL) within 1 week after admission. The levels of serum interleukin-6, tumor necrosis factor-α, interleukin-10, transforming growth factor-β and the percentage of lymphocyte subsets in the peripheral blood were determined in the two groups before and 1, 4 weeks after treatment. The model for end-stage liver disease (MELD) score and Child-Pugh score of 118 patients after treatment for 12 weeks were observed and recorded, and liver failure, complications and survival during follow-up period in the two groups were observed.

RESULTS AND CONCLUSION: After treatment for 1 and 4 weeks, the levels of serum interleukin-6 and tumor necrosis factor-α in the observation group were significantly lower than those in the control group (P < 0.05 or P < 0.001), but the levels of serum interleukin-10 and transforming growth factor-β in the observation group were significantly higher than those in the control group (P < 0.05 or P < 0.001). After treatment for 1 week, the percentages of CD3⁺CD4⁺T cell and CD4⁺CD25⁺Treg cells in the observation group were significantly higher than those in the control group (P < 0.001), but the percentages of CD3⁺CD8⁺ T cells and CD3^-CD19⁺ B cells were significantly lower than those in the control group (P < 0.05 or P < 0.001). After treatment for 4 weeks, the percentages of CD3⁺ T cell ,CD3⁺CD4⁺ T cells and CD4⁺CD25⁺ Treg cells in the observation group were significantly higher than those in the control group (P < 0.05 or P < 0.001), but the percentages of CD3⁺CD8⁺ T cell and CD3^-CD19⁺ B cells were significantly lower than those in the control group (P < 0.05 or P < 0.001). After treatment for 12 weeks, the MELD and Child-Pugh scores in the observation group were significantly lower than those in the control group (P < 0.05). During the follow-up period, none of the cases in the observation group developed liver failure, but five cases in the control group did. In addition, the incidence of complications and cumulative mortality in the observation group were significantly lower than those in the control group (P < 0.05). These results show that the human umbilical cord mesenchymal stem cell transplantation may alleviate liver inflammation and improve liver function, and then may reduce the incidence of hepatic failure and mortality for patients with decompensated cirrhosis due to hepatitis B.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Umbilical Cord, Mesenchymal Stem Cell Transplantation, Liver Cirrhosis, Tissue Engineering

CLC Number:

R394.2

Fang Xue-qing, Zhang Jun-fei, Song Hai-yan, Chen Zhao-lin, Dong Jing, Pan Jin-jin, Chen Xi, Liu Bo, Chen Cong-xin. Effects of human umbilical cord mesenchymal stem cell therapy on the immune function and prognosis in patients with decompensated liver cirrhosis due to hepatitis B[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(17): 2696-2701.

Figures/Tables 1

2.1 参与者数量分析在随访4周时，观察组有1例失访；对照组有1例死亡，2例失访。随访12周时，观察组有2例死亡，2例失访；对照组有3例死亡，3例失访。随访52周时，观察组有8例死亡，2例失访；对照组有15例死亡，4例失访。随访过程中死亡及失访病例均纳入分析，死亡及失访患者均按照无效处理，见图1。 2.2 两组基线情况按年龄、性别、治疗前谷丙转氨酶、总胆红素、白蛋白、凝血酶原时间、Meld评分、Child-Pugh评分将两组患者进行分层比较，差异均无显著性意义(P < 0.05)，见表1。 2.3 两组血清细胞因子水平变化情况治疗前，两组血清白细胞介素6、肿瘤坏死因子α、白细胞介素10、转化生长因子β水平比较差异无显著性意义(P > 0.05)；与对照组相比，观察组治疗后1，4周的血清白细胞介素6、肿瘤坏死因子α下降更明显(P < 0.001)，血清白细胞介素10、转化生长因子β水平上升更显著(P < 0.05)，见表2。 2.4 两组外周血淋巴细胞亚群测定结果治疗前，两组CD3+T细胞、CD3+CD4+T细胞、CD3+CD8+T细胞、CD4+CD25+Treg细胞、CD3-CD16+NK细胞、CD3-CD19+B细胞比例比较差异无显著性意义(P > 0.05)；与对照组相比，观察组治疗后1周的CD3+CD4+T细胞、CD4+CD25+Treg细胞上升更显著(P < 0.001)，CD3+CD8+T细胞和CD3-CD19+B细胞下降更显著(P < 0.001或P < 0.05)；两组CD3+T细胞和CD3-CD16+NK细胞比例差异均无显著性意义(P > 0.05)。与对照组相比，观察组治疗后4周的CD3+T细胞、CD3+CD4+T细胞、Treg细胞上升更显著(P < 0.05或P < 0.001)，CD3+CD8+T细胞和CD3-CD19+B细胞下降更显著(P < 0.001或P < 0.05)；两组CD3-CD16+NK细胞比例比较差异无显著性意义(P > 0.05)，见表3。 2.5 两组患者MELD和Child-Pugh评分变化情况与对照组相比，观察组治疗后12周的MELD和Child-Pugh评分显著下降(P < 0.05)，见表4。 2.6 随访期间两组患者生存情况、发生肝衰竭例数及并发症情况 ①随访期间，观察组死亡及失访15例，对照组死亡及失访28例，死亡原因为肝肾综合征、感染、上消化道大出血、肝性脑病等严重并发症；两组患者1年生存率分别为74.6%(44/59)和52.5%(31/59)，组间比较差异有显著性意义(χ2=4.365，P=0.037)，见图2；②随访52周，观察组未发生肝衰竭，对照组有5例发生肝衰竭，组间比较差异无显著性意义(P=0.057)；③随访52周，观察组消化道出血5例，感染5例，肝性脑病3例，顽固性腹水2例；对照组消化道出血4例，感染12例，肝性脑病5例，顽固性腹水5例，肝肾综合征3例，观察组并发症发生率低于对照组(P < 0.05)。 2.7 不良反应观察组未发生与细胞移植相关的不良反应。"

References

[1] Motawi TM,Atta HM,Sadik NA,et al.The therapeutic effects of bone marrow-derived mesenchymal stem cells and simvastatin in a rat model of liver fibrosis.Cell Biochem Biophys.2014;68(1):111-125.

[2] 孙慧聪,张国尊,郭金波,等.脐带源间充质干细胞移植治疗肝纤维化及肝硬化的相关机制[J].中国组织工程研究,2015,19(41): 6638-6645.

[3] Glennie S, Soeiro I, Dyson PJ, et al. Bone marrow mesenchymal stem cells induce division arrest anergy of activated T Cells.Blood.2005;105(7):2821-2827.

[4] Ghannam S,Pene J,Torcy-Moquet G,et al.Mesenchymal stem cells inhibit human Th17 cell differentiation and function and induce a T regulatory cell phenotype.J Immunol.2010; 185(1):302-312.

[5] 曾芝雨,李东良,方坚,等.脐带间充质干细胞联合骨髓干细胞移植治疗失代偿期肝硬化:1年随访对照[J].中国组织工程研究,2015, 19(10):1533-1538.

[6] 但刚,金静,吴丽娟,等.脐血干细胞移植治疗肝硬化失代偿后T淋巴细胞亚群的观察[J].国际检验医学志,2011,32(20):2319-2320.

[7] Wang L,Han Q,Chen H,et al.Allogeneic bone marrow mesenchymal stem cell transplantation in patients with UDCA-resistant primary biliary cirrhosis.Stem Cells Dev. 2014;23(20):2482-2489.

[8] Herencia C,Almadén Y,Martínez-Moreno JM,et al.Human mesenchymal stromal cell lysates as a novel strategy to recover liver function.Regen Med.2015;10(1):25-38.

[9] Park M,Kim YH,Woo SY,et al.Tonsil-derived mesenchymal stem cells ameliorate CCl4-induced liver fibrosis in mice via autophagy activation.Sci Rep.2015;5:8616.

[10] Sato Y,Araki H,Kato J,et al.Human mesenchymal stem cells xen-ografted directly to rat liver are differentiated into human hepatocytes without fusion.Blood.2005;106(2):756-763.

[11] Volarevic V,Nurkovic J,Arsenijevic N,et al.Concise review:Therapeutic potential of mesenchymal stem cells for the treatment of acute liver failure and cirrhosis.Stem Cells.2014;32(11):2818-2823.

[12] Eom YW,Shim KW,Baik SK.Mesenchymal stem cell therapy for liver fibrosis. Korean J Intern Med.2015;30:580-589.

[13] Di Nicola M,Carlo-stella C,Magni M,et al.Human bone marrow stromal cells supress T- lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli.Blood.2002;99:3838.

[14] Akiyama K,Chen C,Wang D,et al. Mesenchymal-stem-cell- induced immunoregulation involves FAS-ligand/FAS- mediated T cell apoptosis.Cell Stem Cell.2012;10(5):544-555.

[15] Franquesa M,Hoogdujn MJ,Bestard O,et al. Immunomodulatory effect of mesenchymal stem cells on B cells. Front.Immunol.2012;3:212.

[16] Franquesa M,Mensah EK,Huizinga R,et al.Human adipos tissue-derived mesenchymal stem cells abrogate plasmablast formation and induce regulatory B cells independently of T helper cells. Stem cells.2015;33(3):880-891.

[17] Alikarami F, Yari F, Amirizadeh N, et al. The immunosuppressive activity of amniotic membrane mesenchymal stem cells on T lymphocytes.Avicenna J Med Biotechnol.2015;7(3):90-96.

[18] Guo CH,Han LX,Wan MR,et al.Immunomodulatory effect of bone marrow mesenchymal stem cells on T lymphocytes in patients with decompensated liver cirrhosis.Genet Mol Res. 2015;14(2):7039-7046.

[19] Obermajer N,Popp EC,Soeder Y,et al.Conversion of Th17 into IL-17A(neg) regulatory T cells: a novel mechanism in prolonged allograft survival promoted by mesenchymal stem cell-supported minimized immunosuppressive therapy.J Immunol.2014;193(10):4988-4999.

[20] 中华医学会肝病学分会和感染病学分会.慢性乙型肝炎防治指南(2015更新版)[J].中华肝脏病杂志,2015,23(12):888-905.

[21] Amin MA,Sabry D,Rashed LA,et al.Short-term evaluation of autologous transplantation of bone marrow-derived mesenchymal stem cells in patients with cirrhosis: Egyptian study.Clin Transplant.2013;27(4):607-612.

[22] Jang YO,Kim YJ,Baik SK,et al.Histological improvement following administration of autologous bone marrow-derived mesenchymal stem cells for alcoholic cirrhosis: a pilot study. Liver Int.2014;34(1):33-41.

[23] Tang TJ,Kwekkeboom J,Laman JD,et al.The role of intrahepatic immune effector cells in inflammatory liver injury and viral control during chronic hepatitis B infection.J Viral Hepat.2003;10(3):159-167.

[24] Zhou L,Lopes JE,Chong MM,et al.TGF-beta-induced Foxp3 inhibits Th17 cell differentiation by antagonizing ROR gamma function.Nature.2008;455:236-240.

[25] Banas A,Teratani T,Yamamoto Y,et al.Rapid hepatic fate specification of adipose-derived stem cells and their therapeutic potential for liver failure.J Gastroenterol Hepatol. 2009;24(1):70-77.

[26] Chen X,Gan Y,Li W,et al.The interaction between mesenchymal stem cells and steroids during inflammation. Cell Death Dis.2014;5:e1009.

[27] Kharaziha P,Hellström PM,Noorinayer B,et al.Improvement of liver function in liver cirrhosis patients after autologous mesenchymal stem cell injection: a phase I-II clinical trial.Eur J Gastroenterol Hepatol.2009;21(10):1199-1205.

[28] Gholamrezanezhad A,Mirpour S,Baghern M,et al.In vivo tracking of 111 in-oxine labeled mesenchymal stem cells following infusion in patients with advanced cirrhosis.Nucl Med Biol.2011;38(7):961-967.