Removal of blood group A/B antigen in organs by ex vivo and in vivo administration of endo-β-galactosidase (ABase) for ABO-incompatible transplantation
Removal of blood group A/B antigen in organs by ex vivo and in vivo administration of endo-β-galactosidase (ABase) for ABO-incompatible transplantation
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DOI：
10.1016/j.trim.2008.09.007
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发表时间：
2009-01-01
影响因子：
1.5
通讯作者：
Nakao, Akimasa
中科院分区：
文献类型：
--
作者：
Kobayashi, Takaaki;Liu, DaGe;Nakao, Akimasa
背景资料：尽管有效的治疗方法取得了进展，但器官移植中ABO血型不合仍然是抗体介导的排斥反应的高危因素。方法：在E. coli BL-21。用AB酶消化人A/B红细胞（RBC），并在与人血清孵育后进行流式细胞术分析。将纯化的重组AB酶静脉内给予狒狒。在体内给药前和给药后1、4和24 h，从肾脏和肝脏采集活检组织。用冷UW溶液+/-纯化的重组ABase灌注切除的狒狒肾，并在4 ℃下保存。在离体灌注前和灌注后1 h和4 h进行活检。结果：ABase能有效去除人A/B红细胞中82%的A抗原和95%的B抗原，并能有效抑制抗A/B抗体结合和补体激活。ABase在4T下也保持活性。A型狒狒体内输注ABase后，肾小球和肝窦A抗原表达分别下降85%、9%和13%，但无严重不良反应。离体狒狒肾（B型）经ABase灌注和冷藏后，肾小球B抗原表达水平在1h和4h分别降至49%和6%。结论：这种替代方法可能有助于减少抗体清除和抗B细胞免疫抑制，作为ABO血型不合肾、肝和可能的心脏移植的辅助治疗。(C)2008 Elsevier B.V.保留所有权利。
Background: ABO incompatibility in organ transplantation is still a high risk factor for antibody-mediated rejection, despite the progress in effective treatments. We have explored the possibility of using the enzyme to remove the blood type A/B antigen in organs.Methods: Recombinant endo-ss-galactosidase (ABase), which releases A/B antigen, was produced in E. coli BL-21. Human A/B red blood cells (RBC) were digested with ABase, and subjected to flow cytometric analysis after incubation with human sera. Purified recombinant ABase was intravenously administered to a baboon. Biopsies were taken from kidney and liver before and 1, 4 and 24 h after in vivo administration. Excised baboon kidneys were perfused with cold UW solution +/-purified recombinant ABase and preserved at 4 degrees C. Biopsies were taken before and 1 and 4 h after ex vivo perfusion. The change in A/B antigen expression was analyzed by immumohistochemical study.Results: ABase removed 82% of A antigen and 95% of B antigen in human A/B red blood cells, and suppressed anti-A/B antibody binding and complement activation effectively. ABase was also found to remain active at 4 T. In vivo infusion of ABase into a blood type A baboon demonstrated a marked reduction of A antigen expression in the glomeruli of kidney (85% at 1h, 9% at 4 h and 13% at 24 h) and the sinusoids of liver (47% at 1h, 1% at 4 h and 3% at 24 h) without serious adverse effects. After ex vivo perfusion and cold storage of excised baboon kidney (blood type B) with ABase, the expression levels of B antigen in glomeruli were reduced to 49% at 1h and 6% at 4h.Conclusions: This alternative approach might be useful for minimizing antibody removal and anti-B cell immunosuppression as an adjuvant therapy in ABO-incompatible kidney, liver and possibly heart transplantation. (C) 2008 Elsevier B.V. All rights reserved.