ASSESSMENT OF RECOMBINANT ADENOVIRAL VECTORS FOR HEPATIC GENE-THERAPY
ASSESSMENT OF RECOMBINANT ADENOVIRAL VECTORS FOR HEPATIC GENE-THERAPY
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DOI：
10.1089/hum.1993.4.4-403
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发表时间：
1993-08-01
影响因子：
4.2
通讯作者：
WOO, SLC
中科院分区：
文献类型：
--
作者：
LI, QT;KAY, MA;WOO, SLC
重组腺病毒载体最近已被用于在体外和体内将基因转移到许多不同的细胞类型中。携带大肠杆菌β-半乳糖苷酶（β-gal）基因的重组腺病毒载体用于定量病毒直接输注到门静脉后小鼠肝细胞转导的频率。当输注10（10）个腺病毒颗粒时，如通过x-gal染色所确定的，超过95%的肝细胞在体内被转导。转导方案是相对安全的，因为在转导的动物中没有可检测的辅助病毒产生，并且通过该方法转导的肝外细胞非常少。除非使用大量病毒，否则也没有显著肝脏病理学的证据。然而，转导的肝细胞似乎在体内不存在，因为表达β-gal的肝细胞的百分比随时间下降。手术后4个月，0.5-10%的肝细胞在体内含有可检测的β-gal活性。β-gal阳性细胞的变化与腺病毒DNA量的减少相关。因此，目前的重组腺病毒载体可能在急性肝脏疾病的基因治疗中具有临床应用。
Recombinant adenoviral vectors have recently been used to transfer genes into a number of different cell types in vitro and in vivo. A recombinant adenoviral vector bearing the Escherichia coli beta-galactosidase (beta-gal) gene was used to quantitate the frequency of hepatocyte transduction in the mouse after direct viral infusion into the portal vein. When 10(10) adenoviral particles were infused, over 95% of the hepatocytes were transduced in vivo as determined by x-gal staining. The transduction protocol is relatively safe in that there is no detectable helper virus production in transduced animals and that very few extrahepatic cells are transduced by this method. There is also no evidence of significant liver pathology unless substantially greater quantities of virus are used. However, the transduced hepatocytes do not appear to persist in vivo because the percentage of hepatocytes expressing beta-gal declined over time. Four months after the procedure, 0.5-10% of the hepatocytes contain detectable beta-gal activity in vivo. The change in beta-gal-positive cells correlates with decreasing amounts of adenoviral DNA. Thus, current recombinant adenoviral vectors may have clinical applications in gene therapy for acute hepatic disorders.