Modulating the fibrinolytic system of peripheral blood mononuclear cells with adenovirus.
Modulating the fibrinolytic system of peripheral blood mononuclear cells with adenovirus.
复制标题
用腺病毒调节外周血单核细胞的纤溶系统。
DOI：
10.1089/10430340150504055
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Chuang,JL
中科院分区：
文献类型：
--
作者：
Schleef,RR;Olman,MA;Miles,LA;Chuang,JL
利用白细胞进行基因治疗是一种尚未探索的治疗策略，它将组织型纤溶酶原激活物(t-PA)靶向于纤维蛋白和炎症部位。在这项研究中，观察到五种阳离子脂类以剂量依赖的方式增强腺病毒(Ad)介导的人外周血单个核细胞(PBMCs)t-PA的表达，每粒Ad颗粒的脂分子数在1000到15,000之间(效率：LipofetAMINE>；GenePORTER>；Effectene>；SuperFect>；DMRIE-C)。经Ad/t-PA·LipofetAMINE复合体处理的PBMC在4d内t-PA表达增强，表达t-PA的细胞在体外促进了血浆凝块的溶解。功能和免疫学分析表明，Ad·LipofetAMINE感染方案不影响单核细胞在体外的黏附，也不上调促凝血活性、白细胞介素8或肿瘤坏死因子α的表达。这个系统的潜力被证明是一个体内病毒学模型系统，涉及到向腹膜腔内注射脂多糖来诱导炎症反应。将Ad/t-PA感染的大鼠PBMCs注入脂多糖处理的动物的血管系统中，发现局部纤溶活性增加了4倍。这些数据为使用腺病毒将基因转移到PBMC中提供了一个框架。
Gene therapy utilizing leukocytes is an unexplored therapeutic strategy for targeting tissue-type plasminogen activator (t-PA) to fibrin and sites of inflammation. In this study, five cationic lipids were observed to enhance the adenovirus (Ad)-mediated expression of t-PA in human peripheral blood mononuclear cells (PBMCs) in a dose-dependent manner between 1000 and 15,000 lipid molecules per Ad particle (efficiency:LipofectAMINE > GenePORTER > Effectene > SuperFect > DMRIE-C). PBMCs treated with Ad/t-PA • LipofectAMINE complexes displayed elevated t-PA expression over a 4-day period and the t-PA-expressing cells facilitated the lysis of plasma clotsin vitro. Functional and immunologic assays revealed that the Ad • LipofectAMINE infection protocol did not affect monocyte adhesionin vitroor elevate the expression of procoagulant activity, interleukin 8, or tumor necrosis factor α. The potential of this system was documented with anin vivorat model system that involved the injection of lipopolysaccharide into the peritoneal cavity to induce an inflammatory response. Infusion of Ad/t-PA-infected rat PBMCs into the vasculature of lipopolysaccharide-treated animals was found to increase local fibrinolytic activity by 4-fold. These data provide a framework for utilizing adenovirus to transfer genes into PBMCs.