Coupled effects of polybrene and calf serum on the efficiency of retroviral transduction and the stability of retroviral vectors
Coupled effects of polybrene and calf serum on the efficiency of retroviral transduction and the stability of retroviral vectors
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DOI：
10.1089/hum.1997.8.3-285
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发表时间：
1997-02-10
影响因子：
4.2
通讯作者：
Palsson, BO
中科院分区：
文献类型：
--
作者：
Andreadis, S;Palsson, BO
逆转录病毒上清液中聚凝胺（PB）和小牛血清（CS）的相对浓度对逆转录病毒介导的基因转移效率和逆转录病毒载体的稳定性有相当大的影响。 PR 对莫洛尼鼠白血病病毒 (MMuLV) 衍生载体转导效率的影响强烈依赖于 CS：在固定的 CS 浓度下，转导效率显示为 PB 浓度函数的最大值。增加 CS 浓度将该最大值转移到更高的 PB 浓度，但最大值保持不变。因此，存在使基因转移效率最大化的PB和CS浓度的最佳组合：对于1％、2.5％、5％和10％(vol/vol)CS，PB分别为4.4、8.8、13.2和22μg/ml。此外，PB 的存在显着影响逆转录病毒衰变的动力学。在病毒上清液的衰减期间，在没有 PB 的情况下，逆转录病毒活性的丧失并不遵循简单的指数衰减。病毒灭活的动态显示出一个初始阶段，在此期间转导效率保持恒定，然后呈指数衰减。然而，在逆转录病毒载体的衰变期间，如果存在高PB浓度（13.2μg/ml），则最初的延迟消失，并且从一开始就呈指数衰变。目前的结果表明，除了靶细胞表面发生的病毒-ceh相互作用之外，溶液中还可能发生其他物理化学过程，对逆转录病毒活性产生深远影响，因此它们对于基因治疗特别重要。
The relative concentrations of Polybrene (PB) and calf serum (CS) in retroviral supernatant have considerable effects on the efficiency of retrovirus-mediated gene transfer and the stability of retroviral vectors. The effect of PR on the efficiency of transduction of Moloney murine leukemia virus (MMuLV)-derived vectors is strongly dependent on CS: At a fixed CS concentration, the efficiency of transduction shows a maximum as a function of PB concentration. Increasing the CS concentration shifted this maximum to higher PB concentrations, but the value of the maximum remained the same. Therefore, there were optimal combinations of PB and CS concentrations that maximized the efficiency of gene transfer: 4.4, 8.8, 13.2, and 22 mu g/ml of PB for 1%, 2.5%, 5%, and 10% (vol/vol) CS, respectively. Moreover, the presence of PB affected significantly the kinetics of retroviral decay. The loss of retroviral activity did not follow simple exponential decay in the absence of PB during the decay period of the viral supernatant. The dynamics of viral inactivation showed an initial phase during which the transduction efficiency remained constant followed by exponential decay. However, in the presence of high PB concentrations (13.2 mu g/ml) during the decay period of retroviral vectors, the initial delay was lost and the decay was exponential right from the outset. The present results suggest that in addition to virus-ceh interactions that occur on the target cell surface, other physico-chemical processes may occur in solution that have a profound effect on retroviral activity and therefore they are of particular importance for gene therapy.