医学免疫学课件：异种肝移植的研究进展及其所面临的挑战

1、异种肝移植的研究进展及其所面临的挑战23目 录 ONTENTS背景介绍GTKO猪为供体的相关研究术后凝血功能障碍相关研究异位肝移植1C2总结与展望5431背景介绍4异种移植（Xenotransplantation）不同种属个体遗传背景差异大严重的排斥反应5器官移植需求表1 2016年美国器官移植等待名单表2 美国肝脏移植统计UNOS. United Network for Organ Sharing. Http:/. Updated 2016 Accessed December 21, 2016.6临床病例1、美国洛杉矶西代细纳医院1992年野生型大白猪的肝脏胆汁流出存活24h2、美国Mako

2、wka等1995年野生型大白猪的肝脏血浆洗脱法和特异性抗体移出法肝存活20h，受体存活34h抗体介导的超急性排斥反应Makowa, L, et al. The use of a pig liver xenograft for temporary support of a patient with fulminant hepatic failure. Transplantation59.12(1995):1654-9.72GTKO猪为供体的相关研究8GTKO猪灵长类动物体内的抗-1,3-半乳糖的异种反应性天然抗体猪血管内皮细胞上的-1,3-半乳糖抗原-1,3-半乳糖苷转移酶（-1,3-galac

3、tosy transferase, -GT）基因敲除小型猪（GTKO猪）Ekser, B, et al. Genetically-engineered pig-to-baboon liver xenotransplantation: histopathology of xenografts and native organs. Plos One 7.1(2011): e29720.9Figure 1. Time-line in experimental pig liver xenotransplantation in NHPs. a-CD154 = anti-CD154 monoclonal

4、antibodies; ALG,antilymphocyte globulin; ATG, antithymocyte globulin; Aza, azathioprine; Bela, belatacept; BM, bone marrow; Cs, corticosteroids; CsA, cyclosporine;CVF, cobra venom factor; CyP, cyclophosphamide; FK, tacrolimus; Gal abs, extracorporeal anti-Gal antibody adsorption; GTKO, 1,3-galactosy

5、ltransferase gene-knockout; hCD46, expression of the human regulator of complement, hCD46; hCD55, expression of the humanregulator of complement, CD55; HLT, heterotopic liver transplantation; HT, H-transferase; LoCD2b, anti-CD2 monoclonal antibody; MMF,mycophenolate mofetil; OLT, orthotopic liver tr

6、ansplantation; WBI, whole body irradiation; WT, wild-type.Cooper, D. K., et al. Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic. Transplantation 100(2016).10GTKO/hCD46猪Figure 2 (A) Platelet counts after GTKO/hCD46 orthotopic pig liver transplantation in baboons (n = 6) that sur

7、vived from 4 to 7 days. (B) Histopathology of a GTKO/hCD46 orthotopic pig liver graft in a baboon that survived for 6 daysEkser, B, et al. Impact of thrombocytopenia on survival of baboons with genetically modified pig liver transplants: clinical relevance. American Journal of Transplantation 10.2(2

8、010):273-85.11GTKO猪与-氨基乙酸Figure 3. Thrombocytes were lost immediately after reperfusion.Figure 4. Normal liver tissue without any evidence of rejection,haematoxylin and eosin.Kim, K, et al. Up to 9-day survival and control of thrombocytopenia following alpha1,3-galactosyl transferase knockout swine

9、liver xenotransplantation in baboons. Xenotransplantation 19.4(2012):256-64.123术后凝血功能障碍相关研究13供体内皮细胞Burcin Ekser , et al. Potential factors influencing the development of thrombocytopenia and consumptive coagulopathy after genetically modified pig liver xenotransplantation. Transplant International 2

10、5.8(2012):882-96.Figure 5 The expression of CD106 of genetically-engineered pig liver grafts were examined pre-TX, 2h after reperfusion, and at the time of death or euthanasia (days 47). 14受体组织因子的激活Figure 6 TF immunohistochemistry of human and pig TF in donor and recipient livers before, 2 hours aft

11、er, and 48 hours after transplantation.Figure 7 Measurement of endogenous tissue factor activity for recombinant human TFPI, recombinant pig TFPI, and no TFPI control.Ji, H., et al. Pig BMSCs Transfected with Human TFPI Combat Species Incompatibility and Regulate the Human TF Pathway in Vitro and in

12、 a Rodent Model. Cellular Physiology & Biochemistry International Journal of Experimental Cellular Physiology Biochemistry & Pharmacology 36.1(2015):233-49.15SIRP-/CD47 Navarroalvarez, Nalu, and Y. G. Yang. CD47: a new player in phagocytosis and xenograft rejection. Cellular & Molecular Immunology 8

13、.4(2011):285-8.Figure 8(A) SIRP-CD47 corss-species incompatibility.16SIRP-/CD47 Navarroalvarez, Nalu, and Y. G. Yang. CD47: a new player in phagocytosis and xenograft rejection. Cellular & Molecular Immunology 8.4(2011):285-8.Figure 8(B) SIRP-CD47 corss-species incompatibility.17去唾液酸糖蛋白1(Asialoglyco

14、protein receptor 1,ASGR 1)Ahrens, H. E., et al. siRNA mediated knockdown of tissue factor expression in pigs for xenotransplantation. American Journal of Transplantation 15.5(2015):14071414.Figure 9 Potential mechanism of human platelet phagocytosis by pig livers18GGTA1/CMAH基因Butler, J. R., et al. S

15、ilencing Porcine CMAH and GGTA1 Genes Significantly Reduces Xenogeneic Consumption of Human Platelets by Porcine Livers. Transplantation 100.3(2016).Figure 10 A continuous-pass model for the xenogeneic consumption of human platelets by modified porcine livers.194异位肝移植20异位肝移植Cooper, D. K., et al. Pig

16、 Liver Xenotransplantation: A Review of Progress Toward the Clinic. Transplantation 100(2016).Figure 11 Surgical technique of pig left liver lobe transplantation in Tibetanmonkeys.21异位肝移植Cooper, D. K., et al. Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic. Transplantation 100(

17、2016).原位肝移植术切除受体肝脏后原位植入猪肝异位肝移植术优点：不切除受体原肝保留原肝合成凝血因子和抗凝蛋白的功能切除受体脾脏控制术后免疫排异反应224总结与展望23 总结与展望Yang, L., et al. Genome-wide inactivation of porcine endogenous retroviruses (PERVs). Science 350.6264(2015).伦理替代辅助供体 新型转基因猪、嵌合器官供体受体 新型抗凝血、抗排异药物生物安全性 猪逆转录病毒（Porcine endogenous retrovirus, PERV）参考文献1 UNOS. Uni

18、ted Network for Organ Sharing. Http:/. Updated 2016 Accessed December 21, 2016.2 Makowa L, Cramer D V, Hoffman A, et al. The use of a pig liver xenograft for temporary support of a patient with fulminant hepatic failure.J. Transplantation, 1995, 59(12):1654-9.3 Ekser B, Klein E, He J, et al. Genetic

19、ally-engineered pig-to-baboon liver xenotransplantation: histopathology of xenografts and native organs.J. Plos One, 2011, 7(1): e29720. 4 Cooper D K, Dou D F, Tao K S, et al. Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic.J. Transplantation, 2016, 100.5 Ekser B, Long C, Echev

20、erri G J, et al. Impact of thrombocytopenia on survival of baboons with genetically modified pig liver transplants: clinical relevance.J. American Journal of Transplantation, 2010, 10(2):273-85.6 Kim K, Schuetz C, Elias N, et al. Up to 9-day survival and control of thrombocytopenia following alpha1,

21、3-galactosyl transferase knockout swine liver xenotransplantation in baboons.J. Xenotransplantation, 2012, 19(4):256-64.7 Burcin Ekser, Chih C. Lin, Long C, et al. Potential factors influencing the development of thrombocytopenia and consumptive coagulopathy after genetically modified pig liver xeno

22、transplantationJ. Transplant International, 2012, 25(8):882-96.8 Ji H, Li X, Yue S, et al. Pig BMSCs Transfected with Human TFPI Combat Species Incompatibility and Regulate the Human TF Pathway in Vitro and in a Rodent Model.J. Cellular Physiology & Biochemistry International Journal of Experimental Cellular Physiology Biochemistry &