Distribution and expression of porcine endogenous retroviruses in multi-transgenic pigs generated for xenotransplantation
Distribution and expression of porcine endogenous retroviruses in multi-transgenic pigs generated for xenotransplantation
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DOI：
10.1111/j.1399-3089.2009.00515.x
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发表时间：
2009-03-01
影响因子：
3.9
通讯作者：
Denner, Joachim
中科院分区：
文献类型：
--
作者：
Dieckhoff, Britta;Kessler, Barbara;Denner, Joachim
Dieckhoff B，Kessler B，Jobst D，Kues W，Petersen B，Pfeifer A，Kurth R，Niemann H，Wolf E，Denner J. Distribution and expression of porcine endogenous retroviruses in multi-transgenic pigs generated for xenotransplantation.异种移植2009; 16：64-73. (C)2009 John Wiley & Sons A/S.必须筛选用于异种移植的多转基因猪的猪内源性逆转录病毒（PERV）的存在和表达，以选择具有低PERV负荷的动物。转基因猪的产生也可能与邻近或进入PERV原病毒基因座的转基因整合有关，潜在地导致增强的病毒表达。使用聚合酶链反应（PCR）筛选非转基因动物、单转基因和多转基因猪中PERV-A、-B和-C以及重组PERV-A/C的存在。用真实的时间反转录-PCR检测PERV的表达。一项基于有丝分裂原激活外周血单核细胞中PERV的试验被用来区分PERV生产量低和高的动物。181只动物中共有176只（97.2%）在生殖系中携带PERV-C，64只动物中有18只在淋巴细胞基因组中携带PERV-A/C，但在生殖系中未携带。PERV在所有动物中的表达都很低，并且在转基因猪和非转基因动物之间没有差异。PERV的表达在不同的猪品系之间存在差异。在小型猪中发现了最高的表达，并且将其他猪系与小型猪杂交导致后代中PERV表达增加。然而，在所有转基因动物中没有观察到病毒蛋白的表达和颗粒释放。在（多）转基因猪中没有观察到PERV表达升高的证据。PERV表达的差异与动物的遗传背景有关，而与特定的转基因无关。小型猪始终具有最高水平的PERV表达，并且与没有小型猪背景的动物相比，具有小型猪背景的动物具有更高的表达水平。
Dieckhoff B, Kessler B, Jobst D, Kues W, Petersen B, Pfeifer A, Kurth R, Niemann H, Wolf E, Denner J. Distribution and expression of porcine endogenous retroviruses in multi-transgenic pigs generated for xenotransplantation. Xenotransplantation 2009; 16: 64-73. (C) 2009 John Wiley & Sons A/S.Multi-transgenic pigs produced for use in xenotransplantation have to be screened for the presence and expression of porcine endogenous retroviruses (PERV) to select animals with low PERV load. The production of transgenic pigs may also be associated with the integration of the transgene adjacent to or into the locus of a PERV provirus, potentially leading to an enhanced virus expression.Non-transgenic animals, single-transgenic, and multi-transgenic pigs were screened for the presence of PERV-A, -B, and -C and recombinant PERV-A/C using polymerase chain reaction (PCR). PERV expression was determined by real time reverse transcriptase-PCR. An assay based on the activation of PERV in peripheral blood mononuclear cells by mitogens was used to discriminate between low and high PERV producer animals.All animals carried PERV-A and -B. A total of 176 from 181 (97.2%) animals carried PERV-C in the germ line and 18 from 64 animals carried PERV-A/C in the genome of lymphoid cells but not in the germ line. The expression of PERV was very low in all animals and not different between transgenic pigs and non-transgenic animals. PERV expression differed between various pig lines. The highest expression was found in mini-pigs and crossing other pig lines with mini-pigs resulted in increased PERV expression in the progeny. However, expression of viral proteins and particle release were not observed in all transgenic animals.No evidence for elevated PERV expression in (multi-) transgenic pigs was observed. Differences in PERV expression correlated with the genetic background of the animals, not with the specific transgene. Mini-pigs consistently had the highest level of PERV expression and animals with a mini-pig background had a higher level of expression compared with animals without mini-pig background.