Immutol诱导大鼠心脏移植物免疫耐受的研究
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摘要
目的探讨Immutol诱导大鼠心脏移植物产生免疫耐受的作用。方法建立大鼠腹腔异位心脏移植模型。将受体大鼠分成5组:空白对照组(n=6);DMSO组(n=6),使用二甲基亚砜(DMSO)给药至移植物停搏;Immutol组(n=6),使用Immutol给药至移植物停搏;环孢素(CsA)组(n=10),使用CsA给药20 d后停药;联合实验组(n=13),使用Immutol给药60 d后停药,CsA给药20 d后停药。观察各组大鼠心脏移植物的存活时间和病理学变化;检测各组大鼠血清白细胞介素(IL)-10、干扰素(IFN)-γ的含量,以及心脏组织的吲哚胺2,3-双加氧酶(IDO)、纤维蛋白原样蛋白2(Fgl2)的信使核糖核酸(mRNA)表达情况。结果联合实验组心脏移植物可长期存活,存活时间>180 d,均可诱导产生免疫耐受。联合实验组术后39 d心脏移植物的病理评分明显低于CsA组(P<0.05)。联合实验组术后9、39 d血清IL-10和IFN-γ含量均明显高于CsA组(均为P<0.05),联合实验组IL-10和IFN-γ含量随时间延长逐渐升高。联合实验组术后39 d IDO和Fgl2 mRNA表达量明显高于CsA组(均为P<0.05),联合实验组术后IDO的mRNA表达量呈现逐渐升高的趋势。联合实验组术后180 d Fgl2的mRNA表达量明显高于术后9、39 d(均为P<0.05)。结论 Immutol联合CsA可有效抑制急性排斥反应发生,停药后可诱导移植物长期存活,产生免疫耐受。
Objective To investigate the effect of Immutol on inducing the immune tolerance of cardiac grafts in rat models. Methods A rat model of heterotopic abdominal heart transplantation was established. The recipient rats were divided into 5 groups: blank control group(n=6); dimethyl sulfoxide(DMSO) group(n=6), in which DMSO was administered until the cardiac graft arrest; Immutol group(n=6), in which Immutol was administered until the cardiac graft arrest; ciclosporin(CsA) group(n=10), in which CsA was administered for 20 d; combined group(n=13), in which Immutol was given for 60 d combined with CsA for 20 d. The survival time and pathological changes of cardiac grafts in each group were observed. The contents of serum interleukin(IL)-10 and interferon(IFN)-γ were detected. The expression levels of indoleamine 2,3-dioxygenase(IDO) and fibrinogen-like protein 2(Fgl2) messenger RNA(mRNA) in heart tissues of rats in each group were measured. Results In the combined group, the cardiac grafts survived for >180 d and immune tolerance was induced. The pathological score of cardiac grafts in the combined group was significantly lower than that in the CsA group at postoperative 39 d(P<0.05). The levels of serum IL-10 and IFN-γ in the combined group were significantly higher than those in the CsA group at 9 d and 39 d after operation(both P<0.05). The content of serum IL-10 and IFN-γ in the combined group were gradually increased over time. At postoperative 39 d, the expression levels of IDO and Fgl2 mRNA in the combined group were significantly higher than those in the CsA group(both P<0.05).The expression level of IDO mRNA in the combined group tended to gradually elevate after operation. In the combined group, the expression level of Fgl2 mRNA at postoperative 180 d was significantly higher than those at 9 d and 39 d after operation(both P<0.05). Conclusions Combined administration of Immutol and CsA can effectively inhibit the incidence of acute rejection, and maintain the long-term survival of the cardiac grafts and induce immune tolerance after drug withdrawal.
Objective To investigate the effect of Immutol on inducing the immune tolerance of cardiac grafts in rat models. Methods A rat model of heterotopic abdominal heart transplantation was established. The recipient rats were divided into 5 groups: blank control group(n=6); dimethyl sulfoxide(DMSO) group(n=6), in which DMSO was administered until the cardiac graft arrest; Immutol group(n=6), in which Immutol was administered until the cardiac graft arrest; ciclosporin(CsA) group(n=10), in which CsA was administered for 20 d; combined group(n=13), in which Immutol was given for 60 d combined with CsA for 20 d. The survival time and pathological changes of cardiac grafts in each group were observed. The contents of serum interleukin(IL)-10 and interferon(IFN)-γ were detected. The expression levels of indoleamine 2,3-dioxygenase(IDO) and fibrinogen-like protein 2(Fgl2) messenger RNA(mRNA) in heart tissues of rats in each group were measured. Results In the combined group, the cardiac grafts survived for >180 d and immune tolerance was induced. The pathological score of cardiac grafts in the combined group was significantly lower than that in the CsA group at postoperative 39 d(P<0.05). The levels of serum IL-10 and IFN-γ in the combined group were significantly higher than those in the CsA group at 9 d and 39 d after operation(both P<0.05). The content of serum IL-10 and IFN-γ in the combined group were gradually increased over time. At postoperative 39 d, the expression levels of IDO and Fgl2 mRNA in the combined group were significantly higher than those in the CsA group(both P<0.05).The expression level of IDO mRNA in the combined group tended to gradually elevate after operation. In the combined group, the expression level of Fgl2 mRNA at postoperative 180 d was significantly higher than those at 9 d and 39 d after operation(both P<0.05). Conclusions Combined administration of Immutol and CsA can effectively inhibit the incidence of acute rejection, and maintain the long-term survival of the cardiac grafts and induce immune tolerance after drug withdrawal.
引文
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[2]BELLADONNA ML,PUCCETTI P,ORABONA C,et al.Immunosuppression via tryptophan catabolism:the role of kynurenine pathway enzymes[J].Transplantation2007,84(1 Suppl):S17-S20.
[3]TERNESS P,BAUER TM,R?SE L,et al.Inhibition of allogeneic T cell proliferation by indoleamine2,3-dioxygenase-expressing dendritic cells:mediation of suppression by tryptophan metabolites[J].J Exp Med,2002,196(4):447-457.
[4]MUNN DH,SHARMA MD,LEE JR,et al.Potential regulatory function of human dendritic cells expressing indoleamine 2,3-dioxygenase[J].Science,2002,297(5588):1867-1870.
[5]SONG X,ZHANG Y,ZHANG L,et al.Hypoxia enhances indoleamine 2,3-dioxygenase production in dendritic cells[J].Oncotarget,2018,9(14):11572-11580.DOI:10.18632/oncotarget.24098.
[6]WANG Y,MERCHEN TD,FANG X,et al.Regulation of indoleamine 2,3 dioxygenase and its role in a porcine model of acute kidney allograft rejection[J].J Investig Med,2018,66(8):1109-1117.DOI:10.1136/jim-2018-000742.
[7]FALLARINO F,GROHMANN U,VACCA C,et al.Tcell apoptosis by tryptophan catabolism[J].Cell Death Differ,2002,9(10):1069-1077.
[8]FUNESHIMA N,FUJINO M,KITAZAWA Y,et al.Inhibition of allogeneic T-cell responses by dendritic cells expressing transduced indoleamine 2,3-dioxygenase[J].JGene Med,2005,7(5):565-575.
[9]HACKSTEIN H,THOMSON AW.Dendritic cells:emerging pharmacological targets of immunosuppressive drugs[J].Nat Rev Immunol,2004,4(1):24-34.
[10]LI C,QI F,LIU T,et al.Improved cuff technique for establishing a mouse-rat heterotopic cardiac xenotransplantation model[J].Transplant Proc,2015,47(6):2026-2031.DOI:10.1016/j.transproceed.2015.02.028.
[11]SUCHER R,FISCHLER K,OBERHUBER R,et al.IDOand regulatory T cell support are critical for cytotoxic Tlymphocyte-associated Ag-4 Ig-mediated long-term solid organ allograft survival[J].J Immunol,2012,188(1):37-46.DOI:10.4049/jimmunol.1002777.
[12]BIANCHI PKFDC,LEANDRO RM,POSCAI AN,et al.Progesterone decreases in vitro indoleamine2,3-dioxygenase expression in dendritic and CD4+cells from maternal-fetal interface of rats[J].Immunol Invest,2017,46(5):447-459.DOI:10.1080/08820139.2017.1296856.
[13]LI XL,MéNORET S,BEZIE S,et al.Mechanism and localization of CD8 regulatory T cells in a heart transplant model of tolerance[J].J Immunol,2010,185(2):823-833.DOI:10.4049/jimmunol.1000120.
[14]WU Y,YU Z,GONG J,et al.Effects of combined genes of CTLA4Ig and IDO in post-liver transplantation immune tolerance of rats[J].Ann Hepatol,2016,15(5):729-737.DOI:10.5604/16652681.1212524.
[15]HE JG,XIE QL,LI BB,et al.Exosomes derived from IDO1-overexpressing rat bone marrow mesenchymal stem cells promote immunotolerance of cardiac allografts[J].Cell Transplant,2018,12:963689718805375.DOI:10.1177/0963689718805375.
[16]MELLOR AL,LEMOS H,HUANG L.Indoleamine2,3-dioxygenase and tolerance:where are we now?[J].Front Immunol,2017,8:1360.DOI:10.3389/fimmu.2017.01360.
[17]WANG Y,LV S,WANG Q,et al.Mechanisms underlying immune tolerance caused by recombinant Echinococcus granulosus antigens Eg m MDH and Eg10 in dendritic cells[J].PLoS One,2018,13(9):e0204868.DOI:10.1371/journal.pone.0204868.
[18]DANGI A,ZHANG L,ZHANG X,et al.Murine CMVinduces type 1 IFN that impairs differentiation of MDSCs critical for transplantation tolerance[J].Blood Adv,2018,2(6):669-680.DOI:10.1182/bloodadvances.2017012187.
[19]ZHANG L,DEBERGE M,WANG J,et al.Receptor tyrosine kinase MerTK suppresses an allogenic type IIFN response to promote transplant tolerance[J].Am JTransplant,2019,19(3):674-685.DOI:10.1111/ajt.15087.
[20]RO?MAN P,?VAJGER U.The tolerogenic role of IFN-γ[J].Cytokine Growth Factor Rev,2018,41:40-53.DOI:10.1016/j.cytogfr.2018.04.001.
[21]HUA F,CHEN Y,YANG Z,et al.Protective action of bone marrow mesenchymal stem cells in immune tolerance of allogeneic heart transplantation by regulating CD45RB+dendritic cells[J].Clin Transplant,2018,32(4):e13231.DOI:10.1111/ctr.13231.