Absence of PERV infection in baboons after transgenic porcine liver perfusion
Absence of PERV infection in baboons after transgenic porcine liver perfusion
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DOI：
10.1016/j.jss.2004.09.006
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发表时间：
2005-03-01
影响因子：
2.2
通讯作者：
Yamaoka, Y
中科院分区：
文献类型：
--
作者：
Nishitai, R;Ikai, I;Yamaoka, Y
背景异种移植为补充可用于移植的人体器官的短缺提供了很大的希望，但跨物种感染是一个重大问题。特别是猪内源性逆转录病毒（PERV），被认为是对人类构成风险的潜在病原体。我们评估了PERV是否能够在体外猪肝灌注（ECLP）后感染体内非人灵长类动物。从6只人衰变加速因子（h-TNF）转基因仔猪中收获肝脏，并通过门静脉和肝动脉灌注新鲜狒狒血。6只健康狒狒在没有免疫抑制的情况下，用ECLP直接交叉循环13至24小时。定期采集狒狒的外周血和骨髓样本，直至对狒狒实施安乐死，以检查各种器官组织样本。从这些样品中提取基因组DNA，并通过定量PCR检测PERV和猪特异性着丝粒DNA序列。验证表明，该测定法可以在150，000个狒狒细胞的背景中检测一个PERV拷贝，并且在10至10（6）个PERV拷贝的范围内是定量的。在ECLP的初始阶段，在外周血白细胞DNA中检测到大量PERV序列（4.4 × 10（3）-1.6 × 10（4）/1 μ g），但在1周内它们消失。骨髓DNA中含有比外周血长的PERV序列，但PERV信号在1个月内变为阴性。在本研究过程中未观察到PERV DNA复发。猪特异性着丝粒序列也以相同的方式检测。在ECLP后6个月或1年，在从以下器官获得的基因组DNA中未检测到PERV或猪特异性着丝粒序列：皮肤、淋巴结、脾、肝、胰腺、肾、心脏和肺。ECLP没有导致狒狒PERV感染或猪细胞微嵌合体。(c)2004爱思唯尔公司All rights reserved.
Background. Xenotransplantation offers great promise to supplement the shortage of human organs available for transplant, but cross-species infection is a substantial concern. Porcine endogenous retrovirus (PERV), in particular, is thought to pose a risk as a potential pathogen to humans. We evaluated whether PERV is capable of infecting nonhuman primates in vivo after extracorporeal porcine liver perfusion (ECLP).Methods. Livers were harvested from six human decay-accelerating factor (h-DAF) transgenic piglets and perfused with fresh baboon blood via the portal vein and the hepatic artery. Six healthy baboons underwent direct cross-circulation with the ECLP for 13 to 24 h without immunosuppression. Peripheral blood and bone marrow of baboons were sampled periodically until the baboons were euthanized for the examination of various organ tissue samples. Genomic DNA was extracted from those samples and tested for PERV and pig-specific centromeric DNA sequences by quantitative PCR. Validation showed that the assay could detect one copy of PERV in a background of 150,000 baboon cells, and it was quantitative over a range from 10 to 10(6) copies of PERV.Results. PERV sequences were detected in a high number (4.4 X 10(3)-1.6 x 10(4)/1 mu g) in peripheral leukocyte DNA during the initial phases of ECLP, but they disappeared within 1 week. Bone marrow DNA contained PERV sequences longer than peripheral blood, but PERV signals became negative within 1 month. No PERV DNA relapse was seen over the course of this study. Pig-specific centromeric sequences were also detected in the same manner. At 6 months or 1 year after ECLP, no PERV or pig-specific centromeric sequences were detected in the genomic DNA obtained from the following organs: skin, lymph nodes, spleen, liver, pancreas, kidney, heart, and lung.Conclusions. ECLP did not result in PERV infection or pig-cell microchimerism in baboons. (c) 2004 Elsevier Inc. All rights reserved.