受體來源未成熟樹突狀細胞誘導大鼠肝移植免疫低反應性的實驗研究_《中國普外基礎與臨床雜志》

作者：

李立 , 張升寧 , 冉江華 , 劉靜 , 李鑄 , 趙永恒 , 李來邦

昆明市第一人民醫院暨昆明醫學院附屬甘美醫院肝膽胰一科，云南省器官移植研究所肝移植研究中心（昆明650011）;

關鍵詞：

大鼠肝移植排斥反應未成熟樹突狀細胞免疫低反應

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目的研究不負載供者抗原的受者未成熟樹突狀細胞（immature dendritic cell，imDC）對大鼠肝移植的保護作用，探討imDC抑制排斥反應的機理。
方法以SD大鼠作為供體，雄性Wistar大鼠作為受體，隨機分為4組，每組10只，建立肝移植急性排斥反應模型，移植前對受體進行不同的處理。對照組：受體未接受任何處理；環孢素A（CsA）組：術后第2 d開始給予CsA〔10 mg/（kg·d）〕灌胃；成熟樹突狀細胞（mDC）組：受體術前1 d經陰莖背靜脈注射受體來源的mDC 1×106個/只； imDC組：受體術前1 d經陰莖背靜脈注射受體來源的imDC 1×106個/ 只。模型建立后10 d各組隨機處死5只，取移植肝臟組織行HE染色和免疫組化（FasL/Fas）染色，觀察肝臟組織形態結構和排斥反應，根據Banff評分系統判斷排斥反應程度； Western blot分析檢測各組Foxp3編碼的Scurfin蛋白的表達。另留5只大鼠觀察術后存活時間；收集血標本，全自動生化分析儀測定ALT、TBIL；雙抗體夾心ELISA法檢測血清IL-2、IFN-γ、IL-4和IL-10水平。
結果 CsA組和imDC組術后大鼠中位生存時間明顯長于對照組和mDC組，差異有統計學意義（ P ＜ 0.05）。移植術后各組大鼠血清生化檢查：對照組和mDC組ALT及TBIL明顯高于CsA組和imDC組（ P ＜ 0.05）；且IL-2和IFN-γ也明顯高于CsA組和imDC組（ P ＜ 0.01），但IL-4和IL-10明顯低于CsA組和imDC組（ P ＜ 0.01）。移植肝臟形態學檢測：對照組和mDC組與CsA組和 imDC 組比較，排斥反應更嚴重（ P ＜ 0.05）。Western blot檢測： imDC組Foxp3編碼的Scurfin蛋白的表達明顯高于其他3組（ P ＜ 0.05）。
結論 imDC通過阻斷間接識別能明顯延長移植物存活時間。imDC可能通過以下機理誘導免疫低反應：誘導T細胞凋亡；選擇性激活Th2細胞亞群，誘導Th1/Th2偏移；誘導調節性T細胞產生。

引用本文： 李立,張升寧,冉江華,劉靜,李鑄,趙永恒,李來邦. 受體來源未成熟樹突狀細胞誘導大鼠肝移植免疫低反應性的實驗研究. 中國普外基礎與臨床雜志, 2009, 16(1): 32-38. doi: 復制

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2.	李紀鵬，黃怡，王為忠，等. TGF-β1基因修飾的未成熟樹突狀細胞對小腸移植大鼠外周血及移植腸浸潤T細胞的影響 [Ｊ] . 中國普外基礎與臨床雜志， 2008； 15(2)： 96.
3.	Chen-Woan M, Delaney CP, Fournier V, et al . A new protocol for the propagation of dendritic cells from rat bone marrow using recombinant GM-CSF, and their quantification using the mAb OX-62 [Ｊ] . J Immunol Methods， 1995; 178(2)： 157 4 Sun JH, Zeng QH, Wu MC. Experience with orthotopic rat liver transplantation [Ｊ] . Chin Med J (Engl), 1990; 103(2)： 142.
4.	Kamada N, Calne RY. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage [Ｊ] . Transplantation, 1979; 28(1)： 47.
5.	Kashfi A, Mehrabi A, Pahlavan PS, et al . A review of various techniques of orthotopic liver transplantation in the rat [Ｊ] . Transplant Proc, 2005; 37(1)： 185.
6.	張升寧，白建華，劉靜，等. 大鼠原位肝移植模型的手術改進及體會 [Ｊ] . 肝膽外科雜志， 2008； 16(3)： 210.
7.	Pêche H, Trinité B, Martinet B, et al . Prolongation of heart allograft survival by immature dendritic cells generated from recipient type bone marrow progenitors [Ｊ] . Am J Transplant, 2005; 5(2)： 255.
8.	Bériou G, Pêche H, Guillonneau C, et al . Donor-specific allograft tolerance by administration of recipient-derived immature dendritic cells and suboptimal immunosuppression [Ｊ] . Transplantation, 2005; 79(8)： 969.
9.	Demetrius AJ, Batts KP, Dhillon AP, et al . Banff schema for grading liver allograft rejection: An international consensus document [Ｊ] . Hepatology, 1997; 25(3)： 658.
10.	Hübscher S. Diagnosis and grading of liver allograft rejection: a European perspective [Ｊ] . Transplant Proc, 1996; 28(1)： 504.
11.	Hroldt BS, Burattin M, Gunson BK, et al . Does the Banff rejection activity index predict outcome in patients with early acute cellular rejection following liver transplantation? [Ｊ] . Liver Transpl, 2006; 12(7)： 1144.
12.	Pompili M, Mirante VG, Rapaccini GL, et al . Liver transplantation [Ｊ] . Ann Ital Med Int, 2004; 19(1)： 20.
13.	Tiao MM, Lu L, Tao R, et al . Application of recipient-derived dendritic cells to induce donor-specific T-cell hyporesponsiveness [Ｊ] . Transplant Proc, 2004; 36(5)： 1592.
14.	Gould DS, Auchincloss H Jr. Direct and indirect recognition: the role of MHC antigens in graft rejection [Ｊ] . Immunol Today, 1999; 20(2)： 77.
15.	Game DS, Lechler RI. Pathways of allorecognition: implications for transplantation tolerance [Ｊ] . Transpl Immunol, 2002; 10(2-3)： 101.
16.	Benham AM, Sawyer GJ, Fabre JW. Indirect T cell allorecognition of donor antigens contributes to the rejection of vascularized kidney allografts [Ｊ] . Transplantation, 1995; 59(7)： 1028.
17.	Mosmann TR, Sad S. The expanding universe of T-cell subsets: Th1, Th2 and more [Ｊ] . Immunol Today, 1996; 17(3)： 138.
18.	Chen Y, Chen J, Liu Z, et al . Relationship between TH1/TH2 cytokines and immune tolerance in liver transplantation in rats [Ｊ] . Transplant Proc, 2008; 40(8)： 2691.
19.	韓方海，張肇達，李月春，等. 監測Th1/Th2細胞因子變化對豬胰腺移植急性排斥反應早期診斷的意義 [Ｊ] . 四川大學學報（醫學版）， 2006； 37(4)： 587.
20.	Mariotti J, Foley J, Ryan K, et al . Graft rejection as a Th1-type process amenable to regulation by donor Th2-type cells through an interleukin-4/STAT6 pathway [Ｊ] . Blood, 2008; 112(12)： 4765.
21.	Wang YL, Tang ZQ, Gao W, et al . Influence of Th1, Th2, and Th3 cytokines during the early phase after liver transplantation [Ｊ] . Transplant Proc, 2003; 35(8)： 3024.
22.	Gibelli NE, Pinho-Apezzato ML, Miyatani HT, et al . Basiliximab-chimeric anti-IL2-R monoclonal antibody in pediatric liver transplantation: comparative study [Ｊ] . Transplant Proc, 2004; 36(4)： 956.
23.	Pellegrini P, Totaro R, Contasta I, et al . CD30 antigen and multiple sclerosis: CD30, an important costimulatory molecule and marker of a regulatory subpopulation of dendritic cells, is involved in the maintenance of the physiological balance between TH1/TH2 immune responses and tolerance. The role of IFNbeta-1a in the treatment of multiple sclerosis [Ｊ] . Neuroimmunomodulation, 2005; 12(4)： 220.
24.	Min MP, Zhou D, Ichim TE, et al . Inhibitory feedback loop between tolerogenic dendritic cells and regulatory T cells in transplant tolerance [Ｊ] . Immunol, 2003; 170(3)： 1304.
25.	Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3 [Ｊ] . Science, 2003; 299(5609)： 1057.
26.	Albert MH, Liu Y, Anasetti C, et al . Antigen-dependent suppression of alloresponses by Foxp3-induced regulatory T cells in transplantation [Ｊ] . Eur J Immunol, 2005; 35(9)： 2598.
27.	呂凌，張峰，張偉，等. 轉錄因子Foxp3在自發免疫耐受大鼠移植肝內的表達 [Ｊ] . 中國普外基礎與臨床雜志， 2007； 14(2)： 138.
28.	Fontenot JD, Rasmussen JP, Williams LM, et al . Regulatory T cell lineage specification by the forkhead transcription factor foxp3 [Ｊ] . Immunity, 2005; 22(3)： 329.
29.	Wang H, Zhao L, Sun Z, et al . A potential side effect of cyclosporine A: inhibition of CD4+CD25+ regulatory T cells in mice [Ｊ] . Transplantation, 2006; 82 (11)： 1484.

1. Barratt-Boyes SM, Thomson AW. Dendritic cells: tools and targets for transplant tolerance [Ｊ] . Am J Transplant, 2005; 5(12)： 2807.
2. 李紀鵬，黃怡，王為忠，等. TGF-β1基因修飾的未成熟樹突狀細胞對小腸移植大鼠外周血及移植腸浸潤T細胞的影響 [Ｊ] . 中國普外基礎與臨床雜志， 2008； 15(2)： 96.
3. Chen-Woan M, Delaney CP, Fournier V, et al . A new protocol for the propagation of dendritic cells from rat bone marrow using recombinant GM-CSF, and their quantification using the mAb OX-62 [Ｊ] . J Immunol Methods， 1995; 178(2)： 157 4 Sun JH, Zeng QH, Wu MC. Experience with orthotopic rat liver transplantation [Ｊ] . Chin Med J (Engl), 1990; 103(2)： 142.
4. Kamada N, Calne RY. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage [Ｊ] . Transplantation, 1979; 28(1)： 47.
5. Kashfi A, Mehrabi A, Pahlavan PS, et al . A review of various techniques of orthotopic liver transplantation in the rat [Ｊ] . Transplant Proc, 2005; 37(1)： 185.
6. 張升寧，白建華，劉靜，等. 大鼠原位肝移植模型的手術改進及體會 [Ｊ] . 肝膽外科雜志， 2008； 16(3)： 210.
7. Pêche H, Trinité B, Martinet B, et al . Prolongation of heart allograft survival by immature dendritic cells generated from recipient type bone marrow progenitors [Ｊ] . Am J Transplant, 2005; 5(2)： 255.
8. Bériou G, Pêche H, Guillonneau C, et al . Donor-specific allograft tolerance by administration of recipient-derived immature dendritic cells and suboptimal immunosuppression [Ｊ] . Transplantation, 2005; 79(8)： 969.
9. Demetrius AJ, Batts KP, Dhillon AP, et al . Banff schema for grading liver allograft rejection: An international consensus document [Ｊ] . Hepatology, 1997; 25(3)： 658.
10. Hübscher S. Diagnosis and grading of liver allograft rejection: a European perspective [Ｊ] . Transplant Proc, 1996; 28(1)： 504.
11. Hroldt BS, Burattin M, Gunson BK, et al . Does the Banff rejection activity index predict outcome in patients with early acute cellular rejection following liver transplantation? [Ｊ] . Liver Transpl, 2006; 12(7)： 1144.
12. Pompili M, Mirante VG, Rapaccini GL, et al . Liver transplantation [Ｊ] . Ann Ital Med Int, 2004; 19(1)： 20.
13. Tiao MM, Lu L, Tao R, et al . Application of recipient-derived dendritic cells to induce donor-specific T-cell hyporesponsiveness [Ｊ] . Transplant Proc, 2004; 36(5)： 1592.
14. Gould DS, Auchincloss H Jr. Direct and indirect recognition: the role of MHC antigens in graft rejection [Ｊ] . Immunol Today, 1999; 20(2)： 77.
15. Game DS, Lechler RI. Pathways of allorecognition: implications for transplantation tolerance [Ｊ] . Transpl Immunol, 2002; 10(2-3)： 101.
16. Benham AM, Sawyer GJ, Fabre JW. Indirect T cell allorecognition of donor antigens contributes to the rejection of vascularized kidney allografts [Ｊ] . Transplantation, 1995; 59(7)： 1028.
17. Mosmann TR, Sad S. The expanding universe of T-cell subsets: Th1, Th2 and more [Ｊ] . Immunol Today, 1996; 17(3)： 138.
18. Chen Y, Chen J, Liu Z, et al . Relationship between TH1/TH2 cytokines and immune tolerance in liver transplantation in rats [Ｊ] . Transplant Proc, 2008; 40(8)： 2691.
19. 韓方海，張肇達，李月春，等. 監測Th1/Th2細胞因子變化對豬胰腺移植急性排斥反應早期診斷的意義 [Ｊ] . 四川大學學報（醫學版）， 2006； 37(4)： 587.
20. Mariotti J, Foley J, Ryan K, et al . Graft rejection as a Th1-type process amenable to regulation by donor Th2-type cells through an interleukin-4/STAT6 pathway [Ｊ] . Blood, 2008; 112(12)： 4765.
21. Wang YL, Tang ZQ, Gao W, et al . Influence of Th1, Th2, and Th3 cytokines during the early phase after liver transplantation [Ｊ] . Transplant Proc, 2003; 35(8)： 3024.
22. Gibelli NE, Pinho-Apezzato ML, Miyatani HT, et al . Basiliximab-chimeric anti-IL2-R monoclonal antibody in pediatric liver transplantation: comparative study [Ｊ] . Transplant Proc, 2004; 36(4)： 956.
23. Pellegrini P, Totaro R, Contasta I, et al . CD30 antigen and multiple sclerosis: CD30, an important costimulatory molecule and marker of a regulatory subpopulation of dendritic cells, is involved in the maintenance of the physiological balance between TH1/TH2 immune responses and tolerance. The role of IFNbeta-1a in the treatment of multiple sclerosis [Ｊ] . Neuroimmunomodulation, 2005; 12(4)： 220.
24. Min MP, Zhou D, Ichim TE, et al . Inhibitory feedback loop between tolerogenic dendritic cells and regulatory T cells in transplant tolerance [Ｊ] . Immunol, 2003; 170(3)： 1304.
25. Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3 [Ｊ] . Science, 2003; 299(5609)： 1057.
26. Albert MH, Liu Y, Anasetti C, et al . Antigen-dependent suppression of alloresponses by Foxp3-induced regulatory T cells in transplantation [Ｊ] . Eur J Immunol, 2005; 35(9)： 2598.
27. 呂凌，張峰，張偉，等. 轉錄因子Foxp3在自發免疫耐受大鼠移植肝內的表達 [Ｊ] . 中國普外基礎與臨床雜志， 2007； 14(2)： 138.
28. Fontenot JD, Rasmussen JP, Williams LM, et al . Regulatory T cell lineage specification by the forkhead transcription factor foxp3 [Ｊ] . Immunity, 2005; 22(3)： 329.
29. Wang H, Zhao L, Sun Z, et al . A potential side effect of cyclosporine A: inhibition of CD4+CD25+ regulatory T cells in mice [Ｊ] . Transplantation, 2006; 82 (11)： 1484.

《中國普外基礎與臨床雜志》

受體來源未成熟樹突狀細胞誘導大鼠肝移植免疫低反應性的實驗研究

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《中國普外基礎與臨床雜志》

受體來源未成熟樹突狀細胞誘導大鼠肝移植免疫低反應性的實驗研究

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

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