• 昆明市第一人民醫(yī)院暨昆明醫(yī)學(xué)院附屬甘美醫(yī)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）;

目的 研究不負(fù)載供者抗原的受者未成熟樹(shù)突狀細(xì)胞（immature dendritic cell，imDC）對(duì)大鼠肝移植的保護(hù)作用，探討imDC抑制排斥反應(yīng)的機(jī)理。
方法 以SD大鼠作為供體，雄性Wistar大鼠作為受體，隨機(jī)分為4組，每組10只，建立肝移植急性排斥反應(yīng)模型，移植前對(duì)受體進(jìn)行不同的處理。 對(duì)照組： 受體未接受任何處理； 環(huán)孢素A（CsA）組： 術(shù)后第2 d開(kāi)始給予CsA〔10 mg/（kg·d）〕灌胃； 成熟樹(shù)突狀細(xì)胞（mDC）組： 受體術(shù)前1 d經(jīng)陰莖背靜脈注射受體來(lái)源的mDC 1×106個(gè)/只； imDC組： 受體術(shù)前1 d經(jīng)陰莖背靜脈注射受體來(lái)源的imDC 1×106個(gè)/ 只。模型建立后10 d各組隨機(jī)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(tài)結(jié)構(gòu)和排斥反應(yīng)，根據(jù)Banff評(píng)分系統(tǒng)判斷排斥反應(yīng)程度； Western blot分析檢測(cè)各組Foxp3編碼的Scurfin蛋白的表達(dá)。另留5只大鼠觀察術(shù)后存活時(shí)間； 收集血標(biāo)本，全自動(dòng)生化分析儀測(cè)定ALT、TBIL； 雙抗體夾心ELISA法檢測(cè)血清IL-2、IFN-γ、IL-4和IL-10水平。
結(jié)果 CsA組和imDC組術(shù)后大鼠中位生存時(shí)間明顯長(zhǎng)于對(duì)照組和mDC組，差異有統(tǒng)計(jì)學(xué)意義（ P ＜ 0.05）。移植術(shù)后各組大鼠血清生化檢查： 對(duì)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I(xiàn)L-4和IL-10明顯低于CsA組和imDC組（ P ＜ 0.01）。移植肝臟形態(tài)學(xué)檢測(cè)： 對(duì)照組和mDC組與CsA組和 imDC 組比較，排斥反應(yīng)更嚴(yán)重（ P ＜ 0.05）。Western blot檢測(cè)： imDC組Foxp3編碼的Scurfin蛋白的表達(dá)明顯高于其他3組（ P ＜ 0.05）。
結(jié)論 imDC通過(guò)阻斷間接識(shí)別能明顯延長(zhǎng)移植物存活時(shí)間。imDC可能通過(guò)以下機(jī)理誘導(dǎo)免疫低反應(yīng)： 誘導(dǎo)T細(xì)胞凋亡； 選擇性激活Th2細(xì)胞亞群，誘導(dǎo)Th1/Th2偏移； 誘導(dǎo)調(diào)節(jié)性T細(xì)胞產(chǎn)生。

引用本文： 李立,張升寧,冉江華,劉靜,李鑄,趙永恒,李來(lái)邦. 受體來(lái)源未成熟樹(shù)突狀細(xì)胞誘導(dǎo)大鼠肝移植免疫低反應(yīng)性的實(shí)驗(yàn)研究. 中國(guó)普外基礎(chǔ)與臨床雜志, 2009, 16(1): 32-38. doi: 復(fù)制

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(jì)鵬， 黃怡， 王為忠， 等. TGF-β1基因修飾的未成熟樹(shù)突狀細(xì)胞對(duì)小腸移植大鼠外周血及移植腸浸潤(rùn)T細(xì)胞的影響 [Ｊ] . 中國(guó)普外基礎(chǔ)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(shù)改進(jìn)及體會(huì) [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(dá)， 李月春， 等. 監(jiān)測(cè)Th1/Th2細(xì)胞因子變化對(duì)豬胰腺移植急性排斥反應(yīng)早期診斷的意義 [Ｊ] . 四川大學(xué)學(xué)報(bào)（醫(yī)學(xué)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(zhuǎn)錄因子Foxp3在自發(fā)免疫耐受大鼠移植肝內(nèi)的表達(dá) [Ｊ] . 中國(guó)普外基礎(chǔ)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. 1. Barratt-Boyes SM, Thomson AW. Dendritic cells: tools and targets for transplant tolerance [Ｊ] . Am J Transplant, 2005; 5(12)： 2807.
  2. 2. 李紀(jì)鵬， 黃怡， 王為忠， 等. TGF-β1基因修飾的未成熟樹(shù)突狀細(xì)胞對(duì)小腸移植大鼠外周血及移植腸浸潤(rùn)T細(xì)胞的影響 [Ｊ] . 中國(guó)普外基礎(chǔ)與臨床雜志， 2008； 15(2)： 96.
  3. 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. 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. 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. 6. 張升寧， 白建華， 劉 靜， 等. 大鼠原位肝移植模型的手術(shù)改進(jìn)及體會(huì) [Ｊ] . 肝膽外科雜志， 2008； 16(3)： 210.
  7. 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. 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. 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. 10. Hübscher S. Diagnosis and grading of liver allograft rejection: a European perspective [Ｊ] . Transplant Proc, 1996; 28(1)： 504.
  11. 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. 12. Pompili M, Mirante VG, Rapaccini GL, et al . Liver transplantation [Ｊ] . Ann Ital Med Int, 2004; 19(1)： 20.
  13. 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. 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. 15. Game DS, Lechler RI. Pathways of allorecognition: implications for transplantation tolerance [Ｊ] . Transpl Immunol, 2002; 10(2-3)： 101.
  16. 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. 17. Mosmann TR, Sad S. The expanding universe of T-cell subsets: Th1, Th2 and more [Ｊ] . Immunol Today, 1996; 17(3)： 138.
  18. 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. 19. 韓方海， 張肇達(dá)， 李月春， 等. 監(jiān)測(cè)Th1/Th2細(xì)胞因子變化對(duì)豬胰腺移植急性排斥反應(yīng)早期診斷的意義 [Ｊ] . 四川大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）， 2006； 37(4)： 587.
  20. 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. 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. 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. 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. 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. 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. 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. 27. 呂凌， 張峰， 張偉， 等. 轉(zhuǎn)錄因子Foxp3在自發(fā)免疫耐受大鼠移植肝內(nèi)的表達(dá) [Ｊ] . 中國(guó)普外基礎(chǔ)與臨床雜志， 2007； 14(2)： 138.
  28. 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. 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.