PROTAMINE SULFATE AS AN EFFECTIVE ALTERNATIVE TO POLYBRENE IN RETROVIRAL-MEDIATED GENE-TRANSFER - IMPLICATIONS FOR HUMAN-GENE THERAPY
PROTAMINE SULFATE AS AN EFFECTIVE ALTERNATIVE TO POLYBRENE IN RETROVIRAL-MEDIATED GENE-TRANSFER - IMPLICATIONS FOR HUMAN-GENE THERAPY
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DOI：
10.1016/0166-0934(89)90132-8
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发表时间：
1989-02-01
影响因子：
3.1
通讯作者：
ANDERSON, WF
中科院分区：
文献类型：
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作者：
CORNETTA, K;ANDERSON, WF
将聚阳离子硫酸鱼精蛋白与常用的聚凝胺进行比较，以确定其增加逆转录病毒感染效率的能力。使用含有neoR基因和人腺苷脱氨酶（ADA）cDNA的鼠逆转录病毒载体SAX作为细胞感染的标记物。在不同浓度的聚阳离子中在NIH 3 T3细胞上滴定SAX病毒上清液。在5 μ g/ml时观察到鱼精蛋白的最高感染效率，并且比在不存在聚阳离子的情况下进行的感染高7倍。使用鱼精蛋白的感染效率平均为92%。11（SEM）的最高效率获得聚凝胺。当人ADA缺陷型T细胞在鱼精蛋白存在下暴露于SAX上清液时获得的总ADA活性是聚凝胺获得的总ADA活性的83%。小鼠骨髓早期祖细胞（CFU-S）的感染率与每种聚阳离子相似。总之，对于上清液感染，5-10 μ g/ml浓度的鱼精蛋白提供了与聚凝胺基本相同的感染效率，对一系列细胞类型具有低毒性。由于鱼精蛋白被美国食品和药物管理局批准用于人类使用，因此在开发人类基因治疗方案时，它提供了聚凝胺的有效替代品。
The polycation protamine sulfate was compared to polybrene, the usual agent employed, for its ability to increase the efficiency of retroviral infection. The murine retroviral vector SAX, which contains the neoR gene and the human adenosine deaminase (ADA) cDNA, was used as a marker of cell infection. SAX viral supernate was titered on NIH 3T3 cells in varying concentrations of polycation. The highest infection efficiency for protamine was seen at 5.mu.g/ml and was 7-fold greater than infections performed in the absence of polycation. Infection efficiency using protamine avearaged 92% .+-. 11 (SEM) of the highest efficiency obtained with polybrene. Total ADA activity attained when human-ADA deficient T cells were exposed to SAX supernate in the presence of protamine was 83% of that attained with polybrene. The infection rate of mouse bone marrow early progenitor cells (CFU-S) was similar with each polycation. In summary, for supernate infections, concentrations of 5-10 .mu.g/ml of protamine provided essentially the same infection effiency as polybrene with low toxicity on a range of cell types. Since protamine is approved for human use by the U.S. Food and Drug Adminstration it provides as effective alternative to polybrene when developing human gene therapy protocols.