Actions of the Japanese Pancreas and Islet Transplantation Association Regarding Transplanted Human Islets Isolated Using Liberase HI
Actions of the Japanese Pancreas and Islet Transplantation Association Regarding Transplanted Human Islets Isolated Using Liberase HI
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DOI：
10.1016/j.transproceed.2010.09.142
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发表时间：
2010-12-01
影响因子：
0.9
通讯作者：
Teraoka, S.
中科院分区：
文献类型：
--
作者：
Saito, T.;Anazawa, T.;Teraoka, S.
目的.通过认识到Liberase HI分离自在含有脑心浸液肉汤的培养基中生长的溶组织梭菌，表明了将传染性海绵状脑病（TSE）引入胰岛细胞的潜力。日本胰腺和胰岛移植协会的一个国家小组使用Liberase HI在日本实施了一项胰岛移植计划。该计划包括65个来自无心跳供体的胰岛分离和34个移植到18名患者中。在此，我们总结了协会如何长期跟踪这些受助人。我们成立了一个特设委员会，在被告知使用这种酶的相关危险后，对移植了使用Liberase HI分离的胰岛的受体进行跟踪。我们还停止了使用Liberase的胰岛移植。委员会讨论了胶原酶被TSE污染的风险和接受者随访的主要问题。所有受者均接受弥散MRI和EEG检查，然后由Creutzfeldt-Jakob病（CID）专家进行评估和随访。使用表达牛朊病毒蛋白的敲入小鼠进行酶中牛海绵状脑病朊病毒的生物测定。这些检测方法可以检测稀释度为1 × 10（4）的污染朊病毒。在灭活其胶原酶活性后，将Liberase HI注射到基因敲入小鼠的腹腔中。4个月后，通过免疫组织化学染色和免疫印迹法，使用抗朊病毒蛋白抗体对脾匀浆进行检测，评价Liberase 111中朊病毒的感染性。Western印迹和免疫组化染色未检测到Liberase HI中的朊病毒。由CJD专家进行的弥散MRI和EEG评估证实，没有移植受体患有CJD。三年的随访显示，没有一个接受胰岛移植的日本人患上克雅氏病。朊病毒生物测定表明，用于分离供移植的胰岛的Liberase HI不含感染性TSE朊病毒。
Purpose. The potential for introducing transmissible spongiform encephalopathy (TSE) into islet cells was indicated by recognizing that Liberase HI is isolated from Clostridium histolyticum grown in media containing brain heart infusion broth. A national team within the Japanese Pancreas and Islet Transplantation Association implemented an islet transplantation program in Japan using Liberase HI. The program comprised 65 islet isolations from non heart-beating donors and 34 transplants into 18 patients. Herein, we have summarized how the Association followed these recipients over the long term.Procedures. We established an ad hoc committee to follow recipients transplanted with islets isolated using Liberase HI after becoming informed of the associated dangers of using this enzyme. We also stopped islet transplantations using Liberase. The committee addressed the major concerns of the risk of the collagenase being contaminated with TSE and of the recipient follow-up. All recipients were examined by diffusion MRI and EEG and then scheduled for evaluation and follow-up by specialists in Creutzfeldt-Jakob disease (CID). Bioassays of bovine spongiform encephalopathy prions in the enzyme proceeded using knock-in mice expressing bovine prion protein. These assays could detect contaminating prions at a dilution of 1 x 10(4). After inactivating its collagenase activity, Liberase HI was injected into the abdominal cavities of knock-in mice. Four months later, prion infectivity in Liberase 111 was evaluated by immunohistochemical staining and Western blotting of spleen homogenates using anti-prion protein antibodies.Main findings. Western blotting and immunohistochemical staining did not detect prions in Liberase HI. Diffusion MRI and EEG evaluations performed by CJD specialists confirmed that none of the transplanted recipients had CJD.Conclusions. Three years of follow-up revealed that none of the Japanese recipients of islet transplants developed CJD. Prion bioassays showed that the Liberase HI used to isolate islets for transplantation was free of infectious TSE prions.