Liver-directed gene transfer in non-human primates.
Liver-directed gene transfer in non-human primates.
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DOI：
10.1089/hum.1997.8.10-1195
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发表时间：
1997-07
影响因子：
4.2
通讯作者：
Deborah E. Sullivan;S. Dash;Hong Du;Naoki Hiramatsu;Faruk Aydin;Jay K. Kolls;James Blanchard;Gary B. Baskin;Michael A. Gerber
中科院分区：
文献类型：
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作者：
Deborah E. Sullivan;S. Dash;Hong Du;Naoki Hiramatsu;Faruk Aydin;Jay K. Kolls;James Blanchard;Gary B. Baskin;Michael A. Gerber
为了建立一个肝脏定向基因治疗的灵长类动物模型，我们在8只恒河猴（Macaca mulatta）中研究了几种基因转移载体和途径。为此，我们使用第一代，复制缺陷型腺病毒载体携带大肠杆菌lacZ基因（Ad.CMVlacZ）或含有lacZ的质粒（pCMV β）与脂质体转染。将报告基因构建体注入门静脉血管系统、胆总管或隐静脉。腺病毒介导的基因转移通过门静脉导致lacZ的表达在70%以上的肝细胞的第3-7天，但伴随着急性肝炎。腺病毒介导的基因转移通过胆总管导致lacZ表达在不到10%的肝细胞，并伴有门静脉炎症。这些动物产生了显著的免疫应答，如腺病毒抗原诱导的T细胞增殖和中和性抗腺病毒抗体和抗大肠杆菌抗体的产生所证明的。大肠杆菌β-半乳糖苷酶（β-Gal）。免疫应答的激活与第13-21天报告基因的快速减少相关。脂质体介导的基因转移是低效的，没有检测到lacZ在肝脏中的表达。为了限制宿主免疫应答，4只动物用环磷酰胺/泼尼松免疫抑制，然后通过门静脉或隐静脉输注Ad.CMVlacZ。猴子表现出持续表达lacZ长达35天，没有炎症的证据。通过隐静脉转导的灵长类动物在肝脏中显示出与门静脉输注动物相似的β-Gal表达水平。总之，腺病毒介导的基因转移到非人灵长类动物的肝脏通过门静脉或隐静脉是有效的，但它的结果在瞬时表达，并伴随着免疫反应的载体和转基因产品和急性肝炎，而脂质体介导的转移是低效的。宿主免疫反应的操纵可能扩大腺病毒载体用于肝脏定向基因转移的潜在应用。
To develop a primate model for liver-directed gene therapy, we studied several gene transfer vehicles and routes in eight rhesus monkeys (Macaca mulatta). For this purpose, we used first-generation, replication-deficient adenoviral vectors carrying the Escherichia coli lacZ gene (Ad.CMVlacZ) or a lacZ-containing plasmid (pCMV beta) with lipofectamine for transfection. The reporter gene construct was infused into either the portal vasculature, common bile duct, or saphenous vein. Adenovirus-mediated gene transfer via the portal vein resulted in expression of lacZ in over 70% of hepatocytes by days 3-7, but was accompanied by acute hepatitis. Adenovirus-mediated gene transfer via the common bile duct resulted in lacZ expression in less than 10% of hepatocytes and was accompanied by portal inflammation. The animals mounted a significant immune response, as demonstrated by adenoviral antigen-induced T-cell proliferation and production of neutralizing anti-adenovirus antibodies and antibodies to E. coli beta-galactosidase (beta-Gal). Activation of the immune response was associated with rapid decrease of the reporter gene by days 13-21. Lipofectamine-mediated gene transfer was inefficient, and no lacZ expression in the liver was detected. To limit the host immune response, 4 animals were immunosuppressed by cyclophosphamide/prednisone and then infused with the Ad.CMVlacZ via the portal vein or the saphenous vein. The monkeys showed sustained expression of lacZ for up to 35 days with no evidence of inflammation. The primates transduced via the saphenous vein showed a level of beta-Gal expression in the liver similar to that of the portal vein-infused animals. In conclusion, adenovirus-mediated gene transfer to non-human primate livers via the portal vein or saphenous vein is efficient, but it results in transient expression and is accompanied by an immune response to both vector and transgene products and acute hepatitis, whereas lipofectamine-mediated transfer is inefficient. Manipulation of the host immune response may expand potential applications of adenoviral vectors for liver-directed gene transfer.