Kiem Hans-Peter, Allen James, Trobridge Grant, Olson Erik, Keyser Kirsten, Peterson Laura, Russell David W
Clinical Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N D1-100, PO Box 19024, Seattle, WA 98109-1024, USA.
Blood. 2007 Jan 1;109(1):65-70. doi: 10.1182/blood-2006-04-016741. Epub 2006 Sep 12.
Foamy virus (FV) vectors are particularly attractive gene-transfer vectors for stem-cell gene therapy because they form a stable transduction intermediate in quiescent cells and can efficiently transduce hematopoietic stem cells. Here, we studied the use of FV vectors to transduce long-term hematopoietic repopulating cells in the dog, a clinically relevant large animal model. Mobilized canine peripheral blood (PB) CD34+ cells were transduced with an enhanced green fluorescent protein (EGFP)-expressing FV vector in an 18-hour transduction protocol. All 3 dogs studied had rapid neutrophil engraftment to greater than 500/microL with a median of 10 days. Transgene expression was detected in all cell lineages (B cells, T cells, granulocytes, red blood cells, and platelets), indicating multilineage engraftment of transduced cells. Up to 19% of blood cells were EGFP+, and this was confirmed at the DNA level by real-time polymerase chain reaction (PCR) and Southern blot analysis. These transduction rates were higher than the best results we obtained previously with lentiviral vectors in a similar transduction protocol. Integration site analysis also demonstrated polyclonal repopulation and the transduction of multipotential hematopoietic repopulating cells. These data suggest that FV vectors should be useful for stem-cell gene therapy, particularly for applications in which short transduction protocols are critical.
泡沫病毒（FV）载体是干细胞基因治疗中极具吸引力的基因转移载体，因为它们能在静止细胞中形成稳定的转导中间体，并且能够高效转导造血干细胞。在此，我们研究了使用FV载体转导犬类（一种具有临床相关性的大型动物模型）长期造血重建细胞的情况。采用增强型绿色荧光蛋白（EGFP）表达的FV载体，按照18小时的转导方案转导动员的犬外周血（PB）CD34+细胞。所研究的3只犬均迅速出现中性粒细胞植入，数量超过500/微升，植入中位数为10天。在所有细胞谱系（B细胞、T细胞、粒细胞、红细胞和血小板）中均检测到转基因表达，表明转导细胞实现了多谱系植入。高达19%的血细胞为EGFP阳性，这在DNA水平通过实时聚合酶链反应（PCR）和Southern印迹分析得到了证实。这些转导率高于我们先前在类似转导方案中使用慢病毒载体所获得的最佳结果。整合位点分析还证实了多克隆重建以及多能造血重建细胞的转导。这些数据表明，FV载体对于干细胞基因治疗应该是有用的，特别是对于那些短转导方案至关重要的应用。
