Price Mary A, Case Scott S, Carbonaro Denise A, Yu Xiao-Jin, Petersen Denise, Sabo Kathleen M, Curran Michael A, Engel Barbara C, Margarian Hovanes, Abkowitz Janis L, Nolan Garry P, Kohn Donald B, Crooks Gay M
Division of Research Immunology/Bone Marrow Transplantation, Childrens Hospital Los Angeles, Los Angeles, California 90027, USA.
Mol Ther. 2002 Nov;6(5):645-52.
Vectors based on the feline immunodeficiency virus (FIV) have been developed as an alternative to those based on another lentivirus, human immunodeficiency virus-1 (HIV-1), because of theoretical safety advantages. We compared the efficiency of gene transfer and expression in human and feline hematopoietic progenitors using second-generation HIV-1 and FIV-based vectors. Vector pairs were tested using either human cytomegalovirus or murine phospho-glycerate kinase (PGK) internal promoters and were pseudotyped with the vesicular stomatitis virus G protein (VSV-G). Vector proviral copy numbers were similar in human and feline hematopoietic primary cells and cell lines transduced by HIV-1 or FIV vectors, demonstrating that both vectors are able to transfer genes efficiently to these cell types. HIV-1 vectors were well expressed in human primary hematopoietic cells and cell lines. However, transgene expression from FIV vectors was almost undetectable in human hematopoietic cells. In contrast, the FIV vector was expressed well in primary hematopoietic feline cells and human non-hematopoietic cells, demonstrating that low transgene expression from the FIV vector is a phenomenon specific to human hematopoietic cells. Northern blot analysis demonstrated decreased vector transcript levels in human CEM cells transduced with FIV relative to cells transduced with HIV-1, despite high vector copy numbers. No evidence of vector transcript instability was seen in studies of transduced CEM cells treated with actinomycin D. We conclude that FIV vectors can transfer genes into human hematopoietic cells as effectively as HIV-1 vectors, but that unknown elements in the current FIV backbone inhibit expression from FIV vectors in human hematopoietic cells.
由于理论上的安全优势，基于猫免疫缺陷病毒（FIV）的载体已被开发出来，作为基于另一种慢病毒——人类免疫缺陷病毒1型（HIV-1）的载体的替代品。我们使用第二代基于HIV-1和FIV的载体，比较了在人类和猫造血祖细胞中的基因转移和表达效率。使用人类巨细胞病毒或小鼠磷酸甘油酸激酶（PGK）内部启动子对载体对进行测试，并用水泡性口炎病毒G蛋白（VSV-G）进行假型化。HIV-1或FIV载体转导的人类和猫造血原代细胞及细胞系中的载体前病毒拷贝数相似，表明这两种载体都能够有效地将基因转移到这些细胞类型中。HIV-1载体在人类原代造血细胞和细胞系中表达良好。然而，在人类造血细胞中几乎检测不到FIV载体的转基因表达。相反，FIV载体在猫原代造血细胞和人类非造血细胞中表达良好，表明FIV载体转基因表达低是人类造血细胞特有的现象。Northern印迹分析表明，尽管FIV载体拷贝数很高，但与用HIV-1转导的细胞相比，用FIV转导的人类CEM细胞中的载体转录水平降低。在用放线菌素D处理的转导CEM细胞的研究中，未发现载体转录本不稳定的证据。我们得出结论，FIV载体能够像HIV-1载体一样有效地将基因转移到人类造血细胞中，但目前FIV骨架中的未知元件抑制了FIV载体在人类造血细胞中的表达。
