Comparison of the effects of growth factors on retroviral vector-mediated gene transfer and the proliferative status of human hematopoietic progenitor cells.
Comparison of the effects of growth factors on retroviral vector-mediated gene transfer and the proliferative status of human hematopoietic progenitor cells.
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生长因子对逆转录病毒载体介导的基因转移和人类造血祖细胞增殖状态的影响的比较。
作者：
J. Nolta;D. Kohn
我们检测了重组造血生长因子(HGF)、白介素3(IL-3)、白介素6(IL-6)和粒细胞巨噬细胞集落刺激因子(GM-CSF)对逆转录病毒载体介导的人造血祖细胞(HPC)基因转移的促进作用。经GM-CSF或IL-3(而不是IL-6)预培养，以及这三种因子的各自组合均可提高N_2载体向人HPC转导neo基因的效率。IL-3和IL-6的组合产生的G418抗性集落水平(50%-60%)一直显著高于所测试的任何其他组合的HGF。经HGF预培养和氮气转导后，骨髓在长期骨髓培养(LTBMC)中维持2个月。耐G418的HPC水平保持稳定，总HPC含量没有因先前暴露于高刺激性剂量的因子而明显减少。在暴露于肝细胞生长因子后，通过暴露于S时相特异性药物羟基脲而抑制集落形成的百分比来测量HPC的增殖状态。不同HGF促进N2HPC基因转移的能力与它们刺激HPC增殖的程度显著相关。这些结果提示，HGF促进HPC基因转移率的机制是通过刺激细胞增殖。高效率的基因转移技术将有助于研究外源基因在造血细胞中的定量表达，并可能应用于临床基因治疗。
We have examined the ability of the recombinant hematopoietic growth factors (HGF) interleukin-3 (IL-3), IL-6, and granulocyte-macrophage colony-stimulating factor (GM-CSF) to increase retroviral vector-mediated gene transfer into human hematopoietic progenitor cells (HPC). The efficiency of neo gene transfer by the N2 vector into human HPC was enhanced by preculture with either GM-CSF or IL-3 (but not IL-6) and with each combination of the three factors. The combination of IL-3 plus IL-6 consistently produced significantly higher levels of G418-resistant colonies (50-60%) than any of the other combinations of HGF tested. Following preculture with HGF and transduction by N2, marrow was maintained in long-term bone marrow culture (LTBMC) for 2 months. The levels of G418-resistant HPC remained stable, and no apparent depletion of total HPC content resulted from the prior exposure to highly stimulatory doses of factors. The proliferative status of the HPC, following exposure to the HGF, was measured as the percentage of HPC that were inhibited from forming colonies by exposure to the S-phase-specific drug, hydroxyurea. The ability of the different HGF to increase the rate of gene transfer by N2 correlated significantly with the extent to which they stimulated HPC proliferation. These results suggest that the mechanism by which HGF increase rates of gene transfer into HPC is by stimulating cell proliferation. Techniques that produce high rates of gene transfer into long-lived human HPC will facilitate studies to quantitate expression of exogenous genes in hematopoietic cells and may be applicable to clinical gene therapy.
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影响因子：
56.9
作者：
MCCUNE, JM;NAMIKAWA, R;WEISSMAN, IL
通讯作者：
WEISSMAN, IL
DOI：
10.1073/pnas.84.24.9035
发表时间：
1987-12-01
影响因子：
11.1
作者：
IKEBUCHI, K;WONG, GG;OGAWA, M
通讯作者：
OGAWA, M
重组人白细胞介素 3 和粒细胞巨噬细胞集落刺激因子对人单核细胞表现出共同的生物学效应和结合特性。
影响因子：
20.3
作者：
Elliott,MJ;Vadas,MA;Eglinton,JM;Park,LS;To,LB;Cleland,LG;Clark,SC;Lopez,AF
通讯作者：
Lopez,AF
影响因子：
2.7
作者：
Miller Ad;Rosman Gj
通讯作者：
Miller Ad;Rosman Gj
IL-6 和 IL-3 在支持人类造血干细胞增殖方面的协同作用：与 IL-1 α 的比较。
影响因子：
20.3
作者：
Leary,AG;Ikebuchi,K;Hirai,Y;Wong,GG;Yang,YC;Clark,SC;Ogawa,M
通讯作者：
Ogawa,M