Bunnell B A, Kluge K A, Lee-Lin S Q, Byrne E R, Orlic D, Metzger M E, Agricola B A, Wersto R P, Bodine D M, Morgan R A, Donahue R E
Clinical Gene Therapy Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA.
Gene Ther. 1999 Jan;6(1):48-56. doi: 10.1038/sj.gt.3300808.
The transduction efficiencies of immunoselected rhesus macaque (Macaca mulatta) CD34+ cells and colony-forming progenitor cells based on polymerase chain reaction (PCR) analysis were comparable for an amphotropic Moloney murine leukemia virus (MLV) retroviral vector and a retroviral vector derived from the gibbon ape leukemia virus (GaLV) packaging cell line, PG13. On performing autologous transplantation studies using immunoselected CD34+ cells transduced with the GaLV envelope (env) retroviral vector, less than 1% of peripheral blood (PB) contained provirus. This was true whether bone marrow (BM) or cytokine-mobilized PB immunoselected CD34+ cells were reinfused. This level of marking was evident in two animals whose platelet counts never fell below 50,000/microliter and whose leukocyte counts had recovered by days 8 and 10 after having received 1.7 x 10(7) or greater of cytokine-mobilized CD34+ PB cells/kg. Reverse transcriptase(RT)-PCR analysis of CD34+ subsets for both the GaLV and amphotropic receptor were performed. The expression of the GaLV receptor was determined to be restricted to CD34+ Thy-1+ cells, and both CD34+ CD38+ and CD34+ CD38dim cells, while the amphotropic receptor was present on all CD34+ cell subsets examined. Our findings suggest that, in rhesus macaques, PG13-derived retroviral vectors may only be able to transduce a subset of CD34+ cells as only CD34+ Thy-1+ cells express the GaLV receptor.
基于聚合酶链反应（PCR）分析，嗜异性莫洛尼鼠白血病病毒（MLV）逆转录病毒载体和源自长臂猿白血病病毒（GaLV）包装细胞系PG13的逆转录病毒载体对免疫分选的恒河猴（猕猴）CD34+细胞和集落形成祖细胞的转导效率相当。在用GaLV包膜（env）逆转录病毒载体转导的免疫分选CD34+细胞进行自体移植研究时，外周血（PB）中含前病毒的细胞不到1%。无论是回输骨髓（BM）还是细胞因子动员的PB免疫分选CD34+细胞，都是如此。在两只动物中这种标记水平很明显，它们的血小板计数从未低于50,000/微升，并且在接受1.7×10(7)或更多细胞因子动员的CD34+ PB细胞/千克后第8天和第10天白细胞计数已恢复。对GaLV和嗜异性受体的CD34+亚群进行了逆转录酶（RT）-PCR分析。确定GaLV受体的表达仅限于CD34+ Thy-1+细胞以及CD34+ CD38+和CD34+ CD38dim细胞，而在所检测的所有CD34+细胞亚群上都存在嗜异性受体。我们的研究结果表明，在恒河猴中，源自PG13的逆转录病毒载体可能只能转导一部分CD34+细胞，因为只有CD34+ Thy-1+细胞表达GaLV受体。
