CAJ | 학술논문

目的对本研究所构建新型生物人工肝体外支持系统(BAL)应用中,猪内源性逆转录病毒(PERV)的安全性进行验证. 方法正常比格犬接受新型BAL治疗6h,定时收集BAL系统中的血浆及比格犬的全血.提取外周血单核细胞(PBMCs)和血浆中的DNA和RNA进行PCR和RT-PCR检测PERV和猪细胞色素B序列.检测血浆中的逆转录酶(RT)活性以及血浆的体外感染能力.结果比格犬治疗期间均体征平稳并生存至治疗后6个月.仅在BAL内含反应器的循环3中查出PERV RNA和RT活性阳性,包括PBMCs和其他血浆在内的其余样本均未见PERV RNA,PERV DNA和RT活性.在体外感染实验中并无HEK293细胞感染病毒.结论新型BAL并不会导致PERV传播.
목적 대본연구소구건신형생물인공간체외지지계통(BAL)응용중,저내원성역전록병독(PERV)적안전성진행험증. 방법 정상비격견접수신형BAL치료6h,정시수집BAL계통중적혈장급비격견적전혈.제취외주혈단핵세포(PBMCs)화혈장중적DNA화RNA진행PCR화RT-PCR검측PERV화저세포색소B서렬.검측혈장중적역전록매(RT)활성이급혈장적체외감염능력.결과 비격견치료기간균체정평은병생존지치료후6개월.부재BAL내함반응기적순배3중사출PERV RNA화RT활성양성,포괄PBMCs화기타혈장재내적기여양본균미견PERV RNA,PERV DNA화RT활성.재체외감염실험중병무HEK293세포감염병독.결론 신형BAL병불회 도치PERV전파.
Objective To investigate the potential transmissibility of porcine endogenous retrovirus (PERV) from a newly-developed porcine hepatocyte bioartificial liver (BAL) system prior to human clinical trial by using a live canine model.Methods Five normal beagles were treated with the new BAL support system for six hours.Samples of plasma from the BAL system and whole blood from the beagles were collected at regular intervals over the six month study period.DNA and RNA were isohted from both the peripheral blood mononuclear cells (PBMCs) and plasma for evaluation by polymerase chain reaction (PCR) and reverese mnscription (RT)-PCR,respectively,to detect PERV and the Sus scrofa cymchrome B normalization standard.In addition,RT activity and the in vitro infectivity of the plasma were detected in HEK293 cells.Results All five beagles remained in stable physical health throughout the treatment and survived until the end of the study.PERV RNA-positivity and RT activity were only detected in the plasma samples from the 3rd BAL treatment cycle.All other samples,including PBMCs and plasma,were negative for PERV RNA,PERV DNA,and RT activity.In addition,none of the sera samples showed in vitro infectivity.Conclusion Application of our BAL system does not lead to PERV transmission.