Emery D W, Andrews R G, Papayannopoulou T
University of Washington Department of Medicine, Seattle, WA 98195-7720, USA.
Gene Ther. 2000 Mar;7(5):359-67. doi: 10.1038/sj.gt.3301107.
Nonhuman primates are increasingly being used as models for pre-clinical assessment of retrovirus vector expression and function following stem and progenitor cell transduction. We compared the relative susceptibility of CD34+ marrow progenitors from four nonhuman primate species and humans to transduction with amphotropic pseudotyped retrovirus vectors containing the Neo gene. The rate of functional gene transfer was measured by colony formation under G418 selection. Marrow progenitors from pigtail macaques (Macaca nemestrina) were transduced at about twice the rate (19.1 +/- 4.3%) as those from rhesus (11.2 +/- 3.7%) and cynomolgus (7.6 +/- 1.9%) macaques, baboons (7.8 +/- 1.8%), and humans (9.6 +/- 1.7%). Semiquantitative RT/PCR analysis suggests this difference may be due to elevated expression of the amphotropic receptor Pit2 in pigtailed macaque CD34+ cells. Further, transduction rates increased an average 1.6 +/- 0.4-fold when the culture temperature was lowered to 33 degrees C, and 2.1 +/- 0.3-fold when the culture dishes were coated with the fibronectin fragment CH-296. The data presented here point to important differences among nonhuman primate models as well as transduction culture conditions, and suggest that pigtailed macaques may be particularly useful for assessing expression and function of therapeutic retrovirus vectors. Gene Therapy (2000) 7, 359-367.
非人类灵长类动物越来越多地被用作干细胞和祖细胞转导后逆转录病毒载体表达和功能临床前评估的模型。我们比较了来自四种非人类灵长类动物和人类的CD34+骨髓祖细胞对含有Neo基因的双嗜性假型逆转录病毒载体转导的相对敏感性。通过在G418选择下的集落形成来测量功能性基因转移的速率。来自猪尾猕猴(食蟹猴)的骨髓祖细胞的转导率约为恒河猴(11.2 +/- 3.7%)、食蟹猴(7.6 +/- 1.9%)、狒狒(7.8 +/- 1.8%)和人类(9.6 +/- 1.7%)的两倍(19.1 +/- 4.3%)。半定量RT/PCR分析表明,这种差异可能是由于猪尾猕猴CD34+细胞中双嗜性受体Pit2的表达升高所致。此外,当培养温度降至33摄氏度时,转导率平均提高了1.6 +/- 0.4倍,当培养皿用纤连蛋白片段CH-296包被时,转导率提高了2.1 +/- 0.3倍。这里呈现的数据表明非人类灵长类动物模型以及转导培养条件之间存在重要差异,并表明猪尾猕猴可能对评估治疗性逆转录病毒载体的表达和功能特别有用。《基因治疗》(2000年)7卷,359 - 367页。