Garkavenko O, Wynyard S, Nathu D, Simond D, Muzina M, Muzina Z, Scobie L, Hector R D, Croxson M C, Tan P, Elliott B R
Living Cell Technologies Ltd, Manukau 2025, Auckland, New Zealand.
Cell Transplant. 2008;17(12):1381-8. doi: 10.3727/096368908787648056.
Previously a strategy for monitoring of pigs intended for cell transplantation was developed and successfully applied to several representative herds in New Zealand. A designated pathogen-free (DPF) herd has been chosen as a good candidate for xenotransplantation. This herd has previously tested free of infectious agents relevant to xenotransplantation and we present here an in depth study of porcine endogenous retrovirus (PERV) transmission. A panel of assays that describes the constraints for the transmission of PERV has been suggested. It includes a) infectivity test in coculture of DPF pig primary cells with both human and pig target cell lines; b) RT activity in supernatant of stimulated primary cells from DPF pigs; c) viral load in donor's blood plasma; d) PERV proviral copy number in DPF pig genome; e) PERV class C prevalence in the herd and its recombination potential. There was no evidence of PERV transmission from DPF pig tissue to either pig or human cells. Additionally, there was no evidence of PERV RNA present in pig blood plasma. PERV copy number differs in individual pigs from as low as 3 copies to 30 copies and the presence of PERV-C varied between animals and breeds. In all DPF pigs tested, a specific locus for PERV-C potentially associated with the recombination of PERV in miniature swine was absent. Presented data on the PERV transmission allows us to classify the DPF potential donors as "null" or noninfectious pigs.
此前已制定了一种用于监测细胞移植供体猪的策略，并成功应用于新西兰的几个代表性猪群。已选择一个无特定病原体（DPF）猪群作为异种移植的良好候选对象。该猪群此前检测未发现与异种移植相关的感染因子，我们在此展示对猪内源性逆转录病毒（PERV）传播的深入研究。已提出一组描述PERV传播限制的检测方法。它包括：a）DPF猪原代细胞与人和猪靶细胞系共培养的感染性检测；b）DPF猪刺激原代细胞上清液中的RT活性；c）供体血浆中的病毒载量；d）DPF猪基因组中的PERV前病毒拷贝数；e）猪群中C类PERV的流行情况及其重组潜力。没有证据表明PERV从DPF猪组织传播到猪或人细胞。此外，也没有证据表明猪血浆中存在PERV RNA。PERV拷贝数在个体猪中有所不同，低至3拷贝到30拷贝，并且PERV-C的存在在不同动物和品种之间也有所差异。在所有检测的DPF猪中，微型猪中潜在与PERV重组相关的PERV-C特定基因座不存在。所呈现的关于PERV传播的数据使我们能够将DPF潜在供体分类为“无”或无感染性猪。
