Kirkegaard Jeannette S, Ravassard Philippe, Ingvarsen Signe, Diedisheim Marc, Bricout-Neveu Emilie, Grønborg Mads, Frogne Thomas, Scharfmann Raphael, Madsen Ole D, Rescan Claude, Albagli Olivier
J Clin Invest. 2016 Mar 1;126(3):1109-13. doi: 10.1172/JCI83573. Epub 2016 Feb 22.
It has been reported that endogenous retroviruses can contaminate human cell lines that have been passaged as xenotransplants in immunocompromised mice. We previously developed and described 2 human pancreatic β cell lines (EndoC-βH1 and EndoC-βH2) that were generated in this way. Here, we have shown that B10 xenotropic virus 1 (Bxv1), a xenotropic endogenous murine leukemia virus (MuLV), is present in these 2 recently described cell lines. We determined that Bxv1 was also present in SCID mice that were used for in vivo propagation of EndoC-βH1/2 cells, suggesting that contamination occurred during xenotransplantation. EndoC-βH1/2 cells released Bxv1 particles that propagated to human 293T and Mus dunni cells. Mobilization assays demonstrated that Bxv1 transcomplements defective MuLV-based retrovectors. In contrast, common rodent β cell lines, rat INS-1E and RIN-5F cells and mouse MIN6 and βTC3 cells, displayed either no or extremely weak xenotropic helper activity toward MuLV-based retrovectors, although xenotropic retrovirus sequences and transcripts were detected in both mouse cell lines. Bxv1 propagation from EndoC-βH1/2 to 293T cells occurred only under optimized conditions and was overall poorly efficient. Thus, although our data imply that MuLV-based retrovectors should be cautiously used in EndoC-βH1/2 cells, our results indicate that an involuntary propagation of Bxv1 from these cells can be easily avoided with good laboratory practices.
据报道,内源性逆转录病毒可污染在免疫缺陷小鼠中作为异种移植传代的人类细胞系。我们之前开发并描述了通过这种方式产生的2种人类胰腺β细胞系(EndoC-βH1和EndoC-βH2)。在此,我们发现嗜异性内源性鼠白血病病毒(MuLV)——B10嗜异性病毒1(Bxv1)存在于这2种最近描述的细胞系中。我们确定Bxv1也存在于用于EndoC-βH1/2细胞体内传代的SCID小鼠中,这表明污染发生在异种移植过程中。EndoC-βH1/2细胞释放出可传播至人293T细胞和邓氏小鼠细胞的Bxv1颗粒。动员试验表明,Bxv1可对有缺陷的基于MuLV的逆转录病毒载体进行反式互补。相比之下,常见的啮齿动物β细胞系,大鼠INS-1E和RIN-5F细胞以及小鼠MIN6和βTC3细胞,对基于MuLV的逆转录病毒载体要么没有嗜异性辅助活性,要么活性极其微弱,尽管在这两种小鼠细胞系中都检测到了嗜异性逆转录病毒序列和转录本。Bxv1从EndoC-βH1/2传播至293T细胞仅在优化条件下发生,且总体效率低下。因此,尽管我们的数据表明在EndoC-βH1/2细胞中应谨慎使用基于MuLV的逆转录病毒载体,但我们的结果表明,通过良好的实验室操作可以轻松避免Bxv1从这些细胞中意外传播。