Effect of changes in expression of the amphotropic retroviral receptor PiT-2 on transduction efficiency and viral titer: implications for gene therapy.
Effect of changes in expression of the amphotropic retroviral receptor PiT-2 on transduction efficiency and viral titer: implications for gene therapy.
复制标题
双嗜性逆转录病毒受体 PiT-2 表达变化对转导效率和病毒滴度的影响：对基因治疗的影响。
作者：
C. Macdonald;S. Walker;M. Watts;S. Ings;D. Linch;S. Devereux
本研究的目的是量化两性逆转录病毒受体(PIT-2)水平对转导易感性的影响，并确定CD34+造血细胞上发现的PIT-2低水平是否在可能影响基因转移的范围内。将pIT-2基因导入受体缺陷的中国仓鼠卵巢(CHO)细胞，可通过去除四环素诱导其表达。未诱导、完全诱导和部分诱导的转染体中病毒结合测定的PIT-2表达水平与两性逆转录病毒报告载体的转导效率相关。完全诱导的CHO-PIT-2细胞产生了与NIH3T3成纤维细胞相似的明显病毒滴度，而加入四环素后病毒滴度降低了140倍。即使在10 ng/ml浓度的IL-3、IL-6和干细胞因子的预培养下，同一载体制剂与纯化的CD34+外周血干细胞(PBSCs)的结合总是低于未诱导的CHO-PIT-2转染体，这使逆转录病毒结合平均增加了35%。因此，两性逆转录病毒受体PIT-2的表达水平显著限制了在人类基因治疗中重要的靶细胞中观察到的转导效率。
The aim of this study was to quantify the impact of amphotropic retroviral receptor (PiT-2) levels on susceptibility to transduction and to determine whether the low level of PiT-2 found on CD34+ hematopoietic cells is within the range likely to compromise gene transfer. Receptor-deficient Chinese hamster ovary (CHO) cells were transfected with a PiT-2 construct that could be induced by the removal of tetracycline. The level of PiT-2 expression measured by virus binding in uninduced and in fully and partially induced transfectants correlated with the efficiency of transduction by an amphotropic retroviral reporter vector. Fully induced CHO-PiT-2 cells gave apparent viral titers similar to NIH 3T3 fibroblasts while addition of tetracycline reduced titers by up to 140-fold. Binding of the same vector preparation to purified CD34+ peripheral blood stem cells (PBSCs) was always less than to uninduced CHO-PiT-2 transfectants even after preincubation in 10-ng/ml concentrations of IL-3, IL-6, and stem cell factor, which increased retroviral binding by an average of 35%. The level of expression of the amphotropic retroviral receptor PiT-2 thus significantly limits transduction efficiency within the range observed in target cells of importance in human gene therapy.
DOI：
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发表时间：
1990-03
期刊：
Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research
影响因子：
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作者：
O'hara;S. Johann;H. Klinger;D. Blair;H. Rubinson;K. Dunn;P. Sass;S. Vitek;T. Robins
通讯作者：
O'hara;S. Johann;H. Klinger;D. Blair;H. Rubinson;K. Dunn;P. Sass;S. Vitek;T. Robins
影响因子：
3.7
作者：
MARKOWITZ, D;GOFF, S;BANK, A
通讯作者：
BANK, A
生长因子增加双嗜性逆转录病毒与人 CD34+ 骨髓祖细胞的结合。
影响因子：
20.3
作者：
Crooks,GM;Kohn,DB
通讯作者：
Kohn,DB