Adenovirus-mediated transfer of human factor IX gene in immunodeficient and normal mice: evidence for prolonged stability and activity of the transgene in liver.
Adenovirus-mediated transfer of human factor IX gene in immunodeficient and normal mice: evidence for prolonged stability and activity of the transgene in liver.
复制标题
腺病毒介导的免疫缺陷和正常小鼠中人类因子 IX 基因的转移：转基因在肝脏中的长期稳定性和活性的证据。
DOI：
10.1089/vim.1996.9.141
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Kurachi,K
中科院分区：
文献类型：
--
作者：
Yao,SN;Farjo,A;Roessler,BJ;Davidson,BL;Kurachi,K
重组腺病毒最近已成为一种有前途的基因递送载体，可用于治疗各种医学疾病。然而，在体内表达的转基因E1区缺失的腺病毒载体交付是短暂的免疫活性动物。已经提出，宿主免疫机制，特别是细胞毒性T淋巴细胞对腺病毒转导的细胞的破坏可能在限制体内转基因表达的持续时间中起主要作用。在本研究中，从转导的肝组织制备的基因组DNA的Southern印迹分析显示，在免疫活性和免疫缺陷动物中病毒基因组的持续存在，表明腺病毒转导的肝细胞的存活。此外，存活因子IX转基因的活性表达通过在转导的肝组织中存在重组人因子IX以及特异性人因子IX mRNA和蛋白质来显示。人因子IX在正常以及部分免疫缺陷小鼠的循环中的短暂出现主要是由于在这些小鼠中产生小鼠抗人因子IX抗体，而不是宿主对转导细胞的免疫破坏。这些结果表明，在正常动物中，用重组腺病毒载体转导的肝细胞可以逃避宿主免疫机制的破坏。因此，本研究为进一步将腺病毒载体工程改造成用于各种疾病（包括先天性疾病如血友病B）的基因治疗的持久表达系统提供了新的理论基础。
Recombinant adenovirus has recently become a promising gene delivery vehicle that may be used therapeutically for various medical disorders. However,in vivoexpression of transgenes delivered by E1 region-deleted adenoviral vectors is transient in immunocompetent animals. It has been proposed that destruction of adenovirally transduced cells by the host immune mechanisms, particularly cytotoxic T-lymphocytes, may play a major role in limiting the duration of transgene expressionin vivo. In the present study, Southern blot analysis of genomic DNA prepared from transduced liver tissues showed the persistent presence of the viral genome in both immunocompetent and immunodeficient animals, indicating the survival of the adenovirally transduced liver cells. Furthermore, active expression of the surviving factor IX transgenes was shown by the presence of recombinant human factor IX as well as specific human factor IX mRNA and protein in the transduced liver tissues. The transient appearance of human factor IX in the circulation of normal as well as partially immunodeficient mice is primarily due to the generation of mouse antihuman factor IX antibodies in these mice rather than host immune destruction of transduced cells. These results suggest that liver cells transduced with recombinant adenoviral vectors can escape from being destroyed by the host immune mechanism in normal animals. The present study thus provides a new rationale for further engineering of adenoviral vectors into a durable expression system for gene therapy of various diseases including congenital disorders such as hemophilia B.