Department of Molecular Biology, Medical University of Silesia, Narcyzow 1, Sosnowiec, Poland.
Curr Microbiol. 2013 Oct;67(4):505-14. doi: 10.1007/s00284-013-0397-3. Epub 2013 Jun 2.
The pig appears to be the most promising animal donor of organs for use in human recipients. Among several types of pathogens found in pigs, one of the greatest problems is presented by porcine endogenous retroviruses (PERVs). Screening of the source pig herd for PERVs should include analysis of both PERV DNA and RNA. Therefore, the present study focuses on quantitative analysis of PERVs in different organs such as the skin, heart, muscle, and liver and blood of transgenic pigs generated for xenotransplantation. Transgenic pigs were developed to express the human α-galactosidase, the human α-1,2-fucosyltransferase gene, or both genetic modifications of the genome (Lipinski et al., Medycyna Wet 66:316-322, 2010; Lipinski et al., Ann Anim Sci 12:349-356, 2012; Wieczorek et al., Medycyna Wet 67:462-466, 2011). The copy numbers of PERV DNA and RNA were evaluated using real-time Q-PCR and QRT-PCR, respectively. Comparative analysis of all PERV subtypes revealed the following relationships: PERV A > PERV B > PERV C. PERV A and B were found in all samples, whereas PERV C was detected in 47 % of the tested animals. The lowest level of PERV DNA was shown in the muscles for PERV A and B and in blood samples for PERV C. The lowest level of PERV A RNA was found in the skin, whereas those of PERV B and C RNA were found in liver specimens. Quantitative analysis revealed differences in the copy number of PERV subtypes between various organs of transgenic pigs generated for xenotransplantation. Our data support the idea that careful pig selection for organ donation with low PERV copy number may limit the risk of retrovirus transmission to the human recipients.
猪似乎是最有前途的器官供体动物,可用于人类受体。在猪体内发现的几种病原体中,最大的问题之一是内源性猪逆转录病毒(PERVs)。对供体猪群进行 PERV 筛查应包括 PERV DNA 和 RNA 的分析。因此,本研究重点关注用于异种移植的转基因猪的不同器官(如皮肤、心脏、肌肉和肝脏以及血液)中 PERV 的定量分析。转基因猪是为表达人α-半乳糖苷酶、人α-1,2-岩藻糖基转移酶基因或两者的基因组遗传修饰而开发的(Lipinski 等人,Medycyna Wet 66:316-322, 2010;Lipinski 等人,Ann Anim Sci 12:349-356, 2012;Wieczorek 等人,Medycyna Wet 67:462-466, 2011)。使用实时 Q-PCR 和 QRT-PCR 分别评估 PERV DNA 和 RNA 的拷贝数。对所有 PERV 亚型的比较分析表明了以下关系:PERV A > PERV B > PERV C。所有样本中均发现 PERV A 和 B,而在 47%的测试动物中检测到 PERV C。PERV A 和 B 的 PERV DNA 水平在肌肉中最低,而 PERV C 的 PERV DNA 水平在血液样本中最低。PERV A RNA 的最低水平出现在皮肤中,而 PERV B 和 C RNA 的最低水平出现在肝脏标本中。定量分析显示,用于异种移植的转基因猪的不同器官中 PERV 亚型的拷贝数存在差异。我们的数据支持这样一种观点,即仔细选择 PERV 拷贝数低的器官供体可能会限制逆转录病毒向人类受体传播的风险。