• 四川省醫(yī)學(xué)科學(xué)院 四川省人民醫(yī)院肝膽胰外科、細(xì)胞移植中心(四川成都 610072);

目的  研究肝細(xì)胞腹腔移植治療急性肝功能衰竭的免疫排斥反應(yīng)。
方法  用膠原酶灌注法分離幼豬及BALB/c和C57BL/6小鼠肝細(xì)胞;用腹腔注射CCl4的方法建立C57BL/6急性肝功能衰竭模型;將實(shí)驗(yàn)動(dòng)物分為同基因移植組、同種異基因移植組和異種移植組。各組動(dòng)物肝細(xì)胞移植入急性肝功能衰竭小鼠腹腔內(nèi),觀察受體小鼠存活情況,比較各組動(dòng)物T細(xì)胞亞群、免疫球蛋白水平和細(xì)胞因子的變化。
結(jié)果 ?、偈荏w小鼠存活情況:同基因移植組為8/10,同種異基因移植組為6/10,異種移植組為3/10,同基因移植組和同種異基因移植組受體的生存情況明顯好于異種移植組(P<0.05)。②T細(xì)胞亞群變化:移植后7 d內(nèi),同基因移植組外周血中CD4+和CD8+ T細(xì)胞均無(wú)明顯變化,同種異基因移植組CD4+ T細(xì)胞于移植后3 d時(shí)達(dá)高峰(P<0.05),而CD8+ T細(xì)胞7 d內(nèi)無(wú)明顯變化,異種移植組CD4+和CD8+ T細(xì)胞移植后7 d內(nèi)均明顯升高,且于移植后3 d時(shí)達(dá)高峰(P<0.05)。③免疫球蛋白水平:同基因移植組血清免疫球蛋白IgM和IgG水平在移植后0.5、1和3 d時(shí)未見(jiàn)明顯變化,同種異基因移植組和異種移植組的IgM在移植后1 d時(shí)達(dá)高峰(P<0.05),而IgG在移植后3 d時(shí)達(dá)高峰(P<0.05)。④血清細(xì)胞因子水平:移植后7 d,同種異基因移植組和異種移植組的IFN-γ、TNF-α及IL-2血清濃度明顯高于同基因移植組(P<0.05);異種移植組的IL-6血清濃度明顯高于同基因移植組和同種異基因移植組(P<0.05)。
結(jié)論  無(wú)論同種還是異種肝細(xì)胞移植,初期均發(fā)生了強(qiáng)烈的CD4+及CD8+ T細(xì)胞參與的細(xì)胞免疫和較強(qiáng)體液免疫反應(yīng)。

引用本文: 魏玲玲,鄧紹平,黃孝倫. 肝細(xì)胞移植治療急性肝功能衰竭的免疫排斥研究. 中國(guó)普外基礎(chǔ)與臨床雜志, 2012, 19(2): 135-139. doi: 復(fù)制

版權(quán)信息: ?四川大學(xué)華西醫(yī)院華西期刊社《中國(guó)普外基礎(chǔ)與臨床雜志》版權(quán)所有,未經(jīng)授權(quán)不得轉(zhuǎn)載、改編

1. Hughes RD,Mitry RR,Dhawan A.Current status of hepatocyte transplantation[J].Transplantation,2011 Nov 11 [Epub ahead of print]..
2. Ito M,Nagata H,Miyakawa S,et al.Review of hepatocyte transplantation[J].J Hepatobiliary Pancreat Surg,2009,16(2): 97-100..
3. Puppi J,Dhawan A.Human hepatocyte transplantation overview[J].Methods Mol Biol,2009,481:1-16..
4. Akhter J,Johnson LA,Gunasegaram A,et al. Hepatocyte transplantation: a review of laboratory techniques and clinical experiences[J].Surgeon,2007,5(3):155-164..
5. Zern MA.Cell transplantation to replace whole liver transplantation[J].Gastroenterology,2009,136(3):767-769..
6. Li WC,Ralphs KL,Tosh D.Isolation and culture of adult mouse hepatocytes[J].Methods Mol Biol,2010,633:185-196..
7. 王燕,周露婷,章衛(wèi)平,等.高活力原代小鼠肝細(xì)胞的分離與純化[ J].醫(yī)學(xué)研究雜志,2011,40(3):28-31..
8. Song X,Guo Y,Duo S,et al. A mouse model of inducible liver injury caused by tet-on regulated urokinase for studies of hepatocyte transplantation[J].Am J Pathol,2009,175(5):1975-1983..
9. Cho CH,F(xiàn)rancois B,Tilles AW,et al.A new technique for primary hepatocyte expansion in vitro[J].Biotechnol Bioeng, 2008,101(2):345-356..
10. Polson J,Lee WM;American Association for the Study of Liver Disease.AASLD position paper:the management of acute liver failure[J].Hepatology,2005,41(5):1179-1197..
11. Weber A,Groyer-Picard MT,F(xiàn)ranco D,et al.Hepatocyte transplantation in animal models[J].Liver Transpl,2009,15(1):7-14.
12. Seppen J,F(xiàn)ilali EE,Elferink RO.Small animal models of hepatocyte transplantation[J].Methods Mol Biol,2009,481:75-82.
13. Zimmerer JM,Pham TA,Sanders VM,et al.CD8+ T cells negatively regulate IL-4-dependent,IgG1-dominant posttransplant alloantibody production[J].J Immunol,2010,185(12):7285-7292..
14. Tellides G,Pober JS.Interferon-γ axis in graft arteriosclerosis[J].Circ Res,2007,100(5):622-632..
15. Jiang X,Kojo S,Harada M,et al.Mechanism of NKT cell-mediated transplant tolerance[J].Am J Transplant,2007,7(6): 1482-1490..
16. Ekser B,Gridelli B,Veroux M,et al.Clinical pig liver xenotransplantation:how far do we have to go?[J].Xenotransplantation,2011,18(3):158-167..
17. Ekser B,Ezzelarab M,Hara H,et al.Clinical xenotransplantation:the next medical revolution?[J].Lancet,2011 Oct 20[Epub ahead of print]..
18. Bonavita AG,Quaresma K,Cotta-de-Almeida V,et al.Hepatocyte xenotransplantation for treating liver disease[J].Xenotransplantation,2010,17(3):181-187..
19. Bartlett DC,Newsome PN.Xenotransplantation:still a bridge too far?[J].Transplantation,2010,90(5):481-482..
20. Cowan PJ,Robson SC,d’Apice AJ.Controlling coagulation dysregulation in xenotransplantation[J].Curr Opin Organ Transplant,2011,16(2):214-221..
21. Lytras D,Papalois A,Tsaroucha AK,et al.Xenotransplantation of hepatocytes in rats with acute liver failure using sirolimus for immunosuppression[J].J Int Med Res,2010,38(2):546-557.
22. 劉驊,曹暉,吳志勇.不同免疫治療方案誘導(dǎo)大鼠原位肝移植免疫耐受的實(shí)驗(yàn)研究[ J].中國(guó)普通外科雜志, 2009,18(1): 27-30..
23. 陳鐘,蔡鴻宇,湯飛,等.聚乳酸 -O-羧甲基殼聚糖納米粒子對(duì)大鼠腹腔內(nèi)豬肝細(xì)胞異種移植免疫排斥反應(yīng)的影響[ J].中國(guó)組織工程研究與臨床康復(fù),2008,12(32):6287-6291..
24. Bellizzi AM,Legallo RD,Boyd JC,et al.Hepatocyte cytokeratin 7 expression in chronic allograft rejection[J].Am J Clin Pathol,2011,135(2):238-244..
25. Landis CS,Yamanouchi K,Zhou H,et al.Noninvasive evaluation of liver repopulation by transplanted hepatocytes using 31P MRS imaging in mice[J].Hepatology,2006,44(5):12501258..
26. Shimojima N,Shimazu M,Kikuchi H,et al.Serum alcohol dehydrogenase:a sensitive biomarker of ongoing graft function after liver transplantation[J].Clin Transplant,2007,21(4):498-501.
27. Isobe M,F(xiàn)utamatsu H,Suzuki J.Hepatocyte growth factor: Effects on immune-mediated heart diseases[J].Trends Cardiovasc Med,2006,16(6):188-193..
28. Suzuki J,Kawauchi M,Ogawa M,et al.Altered expression of hepatocyte growth factor in cardiac allografts of nonhuman primates[J].Transplant Proc,2006,38(10):3318-3320..
  1. 1. Hughes RD,Mitry RR,Dhawan A.Current status of hepatocyte transplantation[J].Transplantation,2011 Nov 11 [Epub ahead of print]..
  2. 2. Ito M,Nagata H,Miyakawa S,et al.Review of hepatocyte transplantation[J].J Hepatobiliary Pancreat Surg,2009,16(2): 97-100..
  3. 3. Puppi J,Dhawan A.Human hepatocyte transplantation overview[J].Methods Mol Biol,2009,481:1-16..
  4. 4. Akhter J,Johnson LA,Gunasegaram A,et al. Hepatocyte transplantation: a review of laboratory techniques and clinical experiences[J].Surgeon,2007,5(3):155-164..
  5. 5. Zern MA.Cell transplantation to replace whole liver transplantation[J].Gastroenterology,2009,136(3):767-769..
  6. 6. Li WC,Ralphs KL,Tosh D.Isolation and culture of adult mouse hepatocytes[J].Methods Mol Biol,2010,633:185-196..
  7. 7. 王燕,周露婷,章衛(wèi)平,等.高活力原代小鼠肝細(xì)胞的分離與純化[ J].醫(yī)學(xué)研究雜志,2011,40(3):28-31..
  8. 8. Song X,Guo Y,Duo S,et al. A mouse model of inducible liver injury caused by tet-on regulated urokinase for studies of hepatocyte transplantation[J].Am J Pathol,2009,175(5):1975-1983..
  9. 9. Cho CH,F(xiàn)rancois B,Tilles AW,et al.A new technique for primary hepatocyte expansion in vitro[J].Biotechnol Bioeng, 2008,101(2):345-356..
  10. 10. Polson J,Lee WM;American Association for the Study of Liver Disease.AASLD position paper:the management of acute liver failure[J].Hepatology,2005,41(5):1179-1197..
  11. 11. Weber A,Groyer-Picard MT,F(xiàn)ranco D,et al.Hepatocyte transplantation in animal models[J].Liver Transpl,2009,15(1):7-14.
  12. 12. Seppen J,F(xiàn)ilali EE,Elferink RO.Small animal models of hepatocyte transplantation[J].Methods Mol Biol,2009,481:75-82.
  13. 13. Zimmerer JM,Pham TA,Sanders VM,et al.CD8+ T cells negatively regulate IL-4-dependent,IgG1-dominant posttransplant alloantibody production[J].J Immunol,2010,185(12):7285-7292..
  14. 14. Tellides G,Pober JS.Interferon-γ axis in graft arteriosclerosis[J].Circ Res,2007,100(5):622-632..
  15. 15. Jiang X,Kojo S,Harada M,et al.Mechanism of NKT cell-mediated transplant tolerance[J].Am J Transplant,2007,7(6): 1482-1490..
  16. 16. Ekser B,Gridelli B,Veroux M,et al.Clinical pig liver xenotransplantation:how far do we have to go?[J].Xenotransplantation,2011,18(3):158-167..
  17. 17. Ekser B,Ezzelarab M,Hara H,et al.Clinical xenotransplantation:the next medical revolution?[J].Lancet,2011 Oct 20[Epub ahead of print]..
  18. 18. Bonavita AG,Quaresma K,Cotta-de-Almeida V,et al.Hepatocyte xenotransplantation for treating liver disease[J].Xenotransplantation,2010,17(3):181-187..
  19. 19. Bartlett DC,Newsome PN.Xenotransplantation:still a bridge too far?[J].Transplantation,2010,90(5):481-482..
  20. 20. Cowan PJ,Robson SC,d’Apice AJ.Controlling coagulation dysregulation in xenotransplantation[J].Curr Opin Organ Transplant,2011,16(2):214-221..
  21. 21. Lytras D,Papalois A,Tsaroucha AK,et al.Xenotransplantation of hepatocytes in rats with acute liver failure using sirolimus for immunosuppression[J].J Int Med Res,2010,38(2):546-557.
  22. 22. 劉驊,曹暉,吳志勇.不同免疫治療方案誘導(dǎo)大鼠原位肝移植免疫耐受的實(shí)驗(yàn)研究[ J].中國(guó)普通外科雜志, 2009,18(1): 27-30..
  23. 23. 陳鐘,蔡鴻宇,湯飛,等.聚乳酸 -O-羧甲基殼聚糖納米粒子對(duì)大鼠腹腔內(nèi)豬肝細(xì)胞異種移植免疫排斥反應(yīng)的影響[ J].中國(guó)組織工程研究與臨床康復(fù),2008,12(32):6287-6291..
  24. 24. Bellizzi AM,Legallo RD,Boyd JC,et al.Hepatocyte cytokeratin 7 expression in chronic allograft rejection[J].Am J Clin Pathol,2011,135(2):238-244..
  25. 25. Landis CS,Yamanouchi K,Zhou H,et al.Noninvasive evaluation of liver repopulation by transplanted hepatocytes using 31P MRS imaging in mice[J].Hepatology,2006,44(5):12501258..
  26. 26. Shimojima N,Shimazu M,Kikuchi H,et al.Serum alcohol dehydrogenase:a sensitive biomarker of ongoing graft function after liver transplantation[J].Clin Transplant,2007,21(4):498-501.
  27. 27. Isobe M,F(xiàn)utamatsu H,Suzuki J.Hepatocyte growth factor: Effects on immune-mediated heart diseases[J].Trends Cardiovasc Med,2006,16(6):188-193..
  28. 28. Suzuki J,Kawauchi M,Ogawa M,et al.Altered expression of hepatocyte growth factor in cardiac allografts of nonhuman primates[J].Transplant Proc,2006,38(10):3318-3320..