Cancer gene therapy by the in vivo transfer of cytokine-genes in to the artery that leads to tumors with fusogenic liposomes.

我们报道了肿瘤坏死因子（TNF）α在体内通过基因转移在导致肿瘤的动脉中产生时诱导强烈的抗肿瘤免疫反应。我们使用了一个在右足垫携带肉瘤-18O肿瘤的小鼠模型，并将包裹人TNF-α基因的融合脂质体注射到右股动脉中。在这种情况下，仅在通向肿瘤的动脉和肿瘤中检测到人TNF-α。当TNF-α基因被输送到右股动脉时，肿瘤生长显著消退，11只小鼠中有4只完全治愈。当TNF-α基因被递送到左股动脉或肿瘤中或当给予荧光素酶基因时，没有观察到消退。通过注射抗TNF-α、抗CD 4或抗CD 8单克隆抗体抑制肿瘤消退，并且CD 8 + T细胞在TNF-α处理的小鼠的肿瘤中积累。这些结果表明，TNF-α表达水平为1.5%， ...更多信息 局部存在于导致肿瘤的动脉中，通过增强抗肿瘤免疫反应有效地抑制肿瘤生长。此外，中枢神经系统（CNS）基因转移方法的发展是重要的基础研究以及基因治疗的临床研究。阳离子脂质体（CL）是有吸引力的载体，因为它们的安全性和易用性。然而，迄今为止，只有低成功率的报告。我们成功地获得了高转染效率的新生小鼠脑内的CL和细胞质基因表达系统的基础上T7 RNA聚合酶和T7 RNA聚合酶-和酶基因与T7启动子序列。该系统的效率比标准系统高2个数量级，该系统使用CL和荧光素酶基因与劳斯肉瘤病毒启动子pRSVL.In此外，在体外实验中使用LLCMK 2细胞显示，细胞质基因表达发生迅速（6小时内）转染后。而pRSVL诱导荧光素酶表达需要24-48 h。少

We report that tumor necrosis factor (TNF) alpha induced a strong antitumor immune reaction when it was produced in arteries leading to tumors by gene transfer in vivo. We used a mouse model carrying a sarcoma-18O tumor in the right footpad and injected the fusogenic liposomes encapsulating the human TNF-alpha gene into the right femoral artery. Under this condition, human TNF-alpha was detected only in the artery leading to the tumor and in the tumor. There was a significant regression in tumor growth when the TNF-alpha gene was delivered into the right femoral artery, with 4 of 11 mice completely cured. No regression was observed when the TNF-alpha gene was delivered into the left femoral artery or into the tumor or when the luciferase gene was administered. Tumor regression was inhibited by the injection of anti-TNF-alpha, anti-CD4, or anti-CD 8 monoclonal antibody, and CD8+ T cells accumulated in the tumors of TNF-alpha-treated mice. These results suggest that TNF-alpha expressed l … More ocally in the arteries leading to tumors efficiently suppresses tumor growth through reinforcement of an antitumor immune reaction. Additionally, Development of methodologies for gene transfer into the central nervous system (CNS) is important for fundamental research as well as clinical studies for gene therapy. Cationic liposomes (CL) are attractive vectors because of their safety and ease of use. However, to date only low rates of success have been reported. We succeeded in obtaining high transfection efficiencies into the newborn mouse brain in vivo by CL and a cytoplasmic gene expression system based on T7 RNA polymerase and T7 RNA polymerase- and the luciferase-gene with the T7 promoter sequence. This system showed an efficiency rate 2 orders of magnitude higher than the standard system, which used CL and luciferase genes with a Rous sarcoma virus promoter, pRSVL.In addition, in vitro experiments using LLCMK2 cells showed that cytoplasmic gene expression occurred rapidly (within 6 h) after transfection. In contrast, pRSVL required 24-48 h for induction of luciferase expression. Less