Department of Medicine, University of Washington, Seattle, WA 98195, USA.
Hum Gene Ther. 2010 Aug;21(8):1019-27. doi: 10.1089/hum.2010.032.
Abstract The subgroup C feline leukemia virus (FeLV-C) receptor FLVCR is a widely expressed 12-transmembrane domain transporter that exports cytoplasmic heme and is a promising target for retrovirus-mediated gene delivery. Previous studies demonstrated that FeLV-C pseudotype vectors were more efficient at targeting human hematopoietic stem cells than those pseudotyped with gibbon ape leukemia virus (GALV), and thus we developed an all FeLV-C-based packaging system, termed CatPac. CatPac is helper-virus free and can produce higher titer vectors than existing gammaretroviral packaging systems, including systems mixing Moloney murine leukemia virus (MoMLV) Gag-Pol and FeLV-C Env proteins. The vectors can be readily concentrated (>30-fold), refrozen (three to five times), and held on ice (>2 days) with little loss of titer. Furthermore, we demonstrate that CatPac pseudotype vectors efficiently target early CD34(+)CD38(-) stem/progenitor cells, monocytic and erythroid progenitors, activated T cells, mature macrophages, and cancer cell lines, suggesting utility for human cell and cell line transduction and possibly gene therapy.
摘要 亚组 C 猫白血病病毒(FeLV-C)受体 FLVCR 是一种广泛表达的 12 次跨膜域转运蛋白,可输出细胞质血红素,是逆转录病毒介导基因传递的有前途的靶点。先前的研究表明,FeLV-C 假型载体比用长臂猿白血病病毒(GALV)假型化的载体更有效地靶向人类造血干细胞,因此我们开发了一种全 FeLV-C 包装系统,称为 CatPac。CatPac 不含辅助病毒,可产生比现有γ逆转录病毒包装系统更高滴度的载体,包括混合 Moloney 鼠白血病病毒(MoMLV)Gag-Pol 和 FeLV-C Env 蛋白的系统。这些载体可轻松浓缩(>30 倍)、反复冷冻(三到五次)和在冰上保存(>2 天),而滴度损失很小。此外,我们证明 CatPac 假型载体有效地靶向早期 CD34(+)CD38(-)干细胞/祖细胞、单核细胞和红细胞祖细胞、活化的 T 细胞、成熟的巨噬细胞和癌细胞系,这表明其可用于人类细胞和细胞系转导,可能用于基因治疗。