OBJECTIVE Porcine endogenous retroviruses (PERV) involved in pig to human xenotransplantation have raised great concerns because of their ubiquitous nature in pigs and their ability of infecting human cells in vitro. Although no significant cytopathic effect attributed to PERV was evident on PERV-infected human embryonic kidney 293 (HEK293) cells, we did proteomic analysis to investigate the differences of protein profile in order to further characterize the effect of PERV infection. METHODS HEK293 cells were cocultured with porcine peripheral blood mononuclear cells (PBMCs). Protein profiles of PERV-infected and -noninfected HEK293 cells were analyzed by two-dimensional gel electrophoresis (2-DE). Protein spots with at least 1.5-fold alteration were identified by high-definition mass spectrometry (HDMS) analysis. Then real-time RT-PCR and Western blotting were performed to validate the proteomic results. RESULTS Differential analysis of PERV-infected and -noninfected HEK293 cells by 2-DE revealed ten differentially regulated proteins. The proteins identified by HDMS were involved in various cellular pathways including signal transduction, cell apoptosis, and protein synthesis. CONCLUSION The results of this study revealed differentially expressed proteins in HEK293 cells cocultured with porcine PBMCs and implied that these changes were probably induced by PERV infection. These results provide clues and potential links to understanding the molecular effect of the infection by human-tropic PERV.

中文翻译：

---

与猪外周血单个核细胞共培养的人细胞中蛋白质表达的分析。

目的涉及猪到人异种移植的猪内源性逆转录病毒（PERV）引起了人们的极大关注，因为它们在猪中无处不在，并且具有体外感染人细胞的能力。尽管在PERV感染的人类胚胎肾293（HEK293）细胞上没有明显的归因于PERV的明显细胞病变作用，但我们进行了蛋白质组学分析以研究蛋白质谱的差异，以进一步表征PERV感染的作用。方法将HEK293细胞与猪外周血单个核细胞（PBMC）共培养。通过二维凝胶电泳（2-DE）分析了PERV感染和未感染的HEK293细胞的蛋白质谱。通过高清质谱（HDMS）分析鉴定出具有至少1.5倍改变的蛋白质斑点。然后进行实时RT-PCR和蛋白质印迹以验证蛋白质组学结果。结果2-DE对PERV感染的HEK293细胞和未感染的HEK293细胞进行了差异分析，揭示了十种差异调节的蛋白。通过HDMS鉴定的蛋白质参与了各种细胞途径，包括信号转导，细胞凋亡和蛋白质合成。结论这项研究的结果揭示了在与猪PBMC共培养的HEK293细胞中差异表达的蛋白，这暗示这些变化可能是由PERV感染引起的。这些结果提供线索和潜在的联系，以了解由人类型PERV感染的分子效应。结果2-DE对PERV感染的HEK293细胞和未感染的HEK293细胞进行了差异分析，揭示了十种差异调节的蛋白。通过HDMS鉴定的蛋白质参与了各种细胞途径，包括信号转导，细胞凋亡和蛋白质合成。结论这项研究的结果揭示了在与猪PBMC共培养的HEK293细胞中差异表达的蛋白，这暗示这些变化可能是由PERV感染引起的。这些结果提供线索和潜在的联系，以了解由人类型PERV感染的分子效应。结果2-DE对PERV感染的HEK293细胞和未感染的HEK293细胞进行了差异分析，揭示了十种差异调节的蛋白。通过HDMS鉴定的蛋白质参与了各种细胞途径，包括信号转导，细胞凋亡和蛋白质合成。结论这项研究的结果揭示了在与猪PBMC共培养的HEK293细胞中差异表达的蛋白，这暗示这些变化可能是由PERV感染引起的。这些结果提供线索和潜在的联系，以了解由人类型PERV感染的分子效应。结论这项研究的结果揭示了在与猪PBMC共培养的HEK293细胞中差异表达的蛋白，这暗示这些变化可能是由PERV感染引起的。这些结果提供线索和潜在的联系，以了解由人类型PERV感染的分子效应。结论这项研究的结果揭示了在与猪PBMC共培养的HEK293细胞中差异表达的蛋白，这暗示这些变化可能是由PERV感染引起的。这些结果提供线索和潜在的联系，以了解由人类型PERV感染的分子效应。