Le Bas-Bernardet S, Tillou X, Branchereau J, Dilek N, Poirier N, Châtelais M, Charreau B, Minault D, Hervouet J, Renaudin K, Crossan C, Scobie L, Takeuchi Y, Diswall M, Breimer M E, Klar N, Daha M R, Simioni P, Robson S C, Nottle M B, Salvaris E J, Cowan P J, d'Apice A J F, Sachs D H, Yamada K, Lagutina I, Duchi R, Perota A, Lazzari G, Galli C, Cozzi E, Soulillou J-P, Vanhove B, Blancho G
Institut de Transplantation- Urologie- Néphrologie (ITUN), Institut National de la Santé et de la Recherche Médicale (INSERM) UMR-S 1064, Centre Hospitalier Universitaire (CHU) de Nantes, Université de Nantes, Nantes, France.
Am J Transplant. 2015 Feb;15(2):358-70. doi: 10.1111/ajt.12988.
Galactosyl-transferase KO (GalT-KO) pigs represent a potential solution to xenograft rejection, particularly in the context of additional genetic modifications. We have performed life supporting kidney xenotransplantation into baboons utilizing GalT-KO pigs transgenic for human CD55/CD59/CD39/HT. Baboons received tacrolimus, mycophenolate mofetil, corticosteroids and recombinant human C1 inhibitor combined with cyclophosphamide or bortezomib with or without 2-3 plasma exchanges. One baboon received a control GalT-KO xenograft with the latter immunosuppression. All immunosuppressed baboons rejected the xenografts between days 9 and 15 with signs of acute humoral rejection, in contrast to untreated controls (n = 2) that lost their grafts on days 3 and 4. Immunofluorescence analyses showed deposition of IgM, C3, C5b-9 in rejected grafts, without C4d staining, indicating classical complement pathway blockade but alternate pathway activation. Moreover, rejected organs exhibited predominantly monocyte/macrophage infiltration with minimal lymphocyte representation. None of the recipients showed any signs of porcine endogenous retrovirus transmission but some showed evidence of porcine cytomegalovirus (PCMV) replication within the xenografts. Our work indicates that the addition of bortezomib and plasma exchange to the immunosuppressive regimen did not significantly prolong the survival of multi-transgenic GalT-KO renal xenografts. Non-Gal antibodies, the alternative complement pathway, innate mechanisms with monocyte activation and PCMV replication may have contributed to rejection.
半乳糖基转移酶基因敲除(GalT-KO)猪是异种移植排斥问题的一种潜在解决方案,特别是在进行额外基因改造的情况下。我们利用转人CD55/CD59/CD39/HT基因的GalT-KO猪对狒狒进行了维持生命的肾异种移植。狒狒接受了他克莫司、霉酚酸酯、皮质类固醇以及重组人C1抑制剂,并联合使用环磷酰胺或硼替佐米,同时进行或不进行2-3次血浆置换。一只狒狒接受了采用后一种免疫抑制方案的对照GalT-KO异种移植肾。与未治疗的对照狒狒(n = 2)在第3天和第4天失去移植肾不同,所有接受免疫抑制的狒狒在第9天至第15天之间排斥了异种移植肾,出现急性体液排斥的迹象。免疫荧光分析显示,在被排斥的移植肾中有IgM、C3、C5b-9沉积,但无C4d染色,表明经典补体途径被阻断但替代途径被激活。此外,被排斥的器官主要表现为单核细胞/巨噬细胞浸润,淋巴细胞浸润极少。所有受体均未表现出任何猪内源性逆转录病毒传播的迹象,但部分受体显示出异种移植肾内有猪巨细胞病毒(PCMV)复制的证据。我们的研究表明,在免疫抑制方案中添加硼替佐米和血浆置换并不能显著延长多转基因GalT-KO肾异种移植的存活时间。非Gal抗体、替代补体途径、单核细胞激活的固有机制以及PCMV复制可能都参与了排斥反应。