Selective transduction of murine myelomonocytic leukemia cells (WEHI-3B) with regular and RGD-adenoviral vectors.
Selective transduction of murine myelomonocytic leukemia cells (WEHI-3B) with regular and RGD-adenoviral vectors.
复制标题
使用常规载体和 RGD 腺病毒载体选择性转导小鼠骨髓单核细胞白血病细胞 (WEHI-3B)。
DOI：
10.1006/mthe.2000.0221
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Bueren,JA
中科院分区：
文献类型：
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作者：
García-Castro,J;Segovia,JC;García-Sánchez,F;Lillo,R;Gómez-Navarro,J;Curiel,DT;Bueren,JA
在正常骨髓单核细胞对腺病毒载体敏感的基础上，我们研究了常规（Reg-Ad）和基因修饰（RGD-Ad）腺病毒载体选择性转导小鼠骨髓单核细胞白血病细胞（WEHI-3B）的可能性。用100 pfu的Reg-Ad载体/细胞孵育WEHI-3B细胞8小时，导致整个群体对转基因表达呈阳性。在相同的感染条件下，20-30%的小鼠骨髓（BM）细胞对转基因呈阳性。当使用RGD-Ad载体时，WEHI-3B细胞短暂暴露于150 pfu病毒/细胞（10分钟）足以使100%的白血病细胞对标记转基因（EGFP）呈阳性。在这些条件下，只有15-20%的BM细胞和原始造血祖细胞（Lin-Sca-1+细胞）变成EGFP+，表明载体对白血病细胞的选择性提高。用可溶性纤维蛋白（FP）孵育WEHI-3B而不是正常BM细胞抑制Reg-Ad的感染。RGD-Ad的使用绕过了用Reg-Ad转导WEHI-3B细胞所需的FP-CAR相互作用，这表明该要求的废除解释了这些白血病细胞的感染性的改善以及RGD-Ad在靶向WEHI-3B白血病细胞中的选择性。
On the basis of the susceptibility of normal myelomonocytic cells to adenoviral vectors, we have studied the possibility of selectively transducing myelomonocytic murine leukemic cells (WEHI-3B) with regular (Reg-Ad) and genetically modified (RGD-Ad) adenoviral vectors. An 8-h incubation of WEHI-3B cells with 100 pfu of Reg-Ad vectors/cell resulted in the whole population becoming positive for transgene expression. Under identical conditions of infection, 20–30% of mouse bone marrow (BM) cells were positive for the transgene. When RGD-Ad vectors were used, a brief exposure (10 min) of WEHI-3B cells to 150 pfu of the virus/cell was enough for 100% of the leukemia cells to become positive for the marker transgene (EGFP). Under these conditions, only 15–20% of BM cells and of primitive hematopoietic progenitors (Lin−Sca-1+cells) became EGFP+, indicating an improved selectivity of the vectors for the leukemic cells. The incubation of WEHI-3B but not normal BM cells with soluble fiber protein (FP) inhibited the infection with Reg-Ad. The use of the RGD-Ad bypassed the FP-CAR interaction required for the transduction of WEHI-3B cells with Reg-Ad, suggesting that the abrogation of this requirement accounts for the improved infectivity of these leukemic cells and for the selectivity of RGD-Ad in targeting WEHI-3B leukemia cells.