Yang Haiguang, Joo Kye-Il, Ziegler Leslie, Wang Pin
Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, 925 Bloom Walk, Los Angeles, CA 90089, USA.
Pharm Res. 2009 Jun;26(6):1432-45. doi: 10.1007/s11095-009-9853-y. Epub 2009 Mar 4.
The purpose of this study was to investigate the potential of a T-cell-related targeting method using a lentiviral vector-based gene delivery system.
A lentiviral vector system was constructed by co-incorporating an anti-CD3 antibody (OKT3) and a fusogen into individual viral particles. The incorporation of OKT3 and fusogen was analyzed using confocal microscopy and the in vitro transduction efficiency was evaluated using flow cytometry. Blocking reagents (ammonium chloride (NH(4)Cl) and soluble OKT3 antibody) were added into vector supernatants during transduction to study the mechanism of this two-molecule targeting strategy. To demonstrate the ability of targeted transduction in vivo, Jurkat.CD3 cells were xenografted subcutaneously into the right flank of each mouse and the lentiviral vector was injected subcutaneously on both sides of each mouse 8 h post-injection. Subsequently, the reporter gene (firefly luciferase) expression was monitored using a noninvasive bioluminescence imaging system.
By co-displaying OKT3 and fusogen on the single lentiviral surface, we could achieve targeted delivery of genes to CD3-positive T-cells both in vitro and in vivo.
These results suggest the potential utility of this engineered lentiviral system as a new tool for cell type-directed gene delivery.
本研究旨在探讨使用基于慢病毒载体的基因递送系统进行T细胞相关靶向方法的潜力。
通过将抗CD3抗体（OKT3）和融合素共掺入单个病毒颗粒构建慢病毒载体系统。使用共聚焦显微镜分析OKT3和融合素的掺入情况，并使用流式细胞术评估体外转导效率。在转导过程中将阻断试剂（氯化铵（NH₄Cl）和可溶性OKT3抗体）添加到载体上清液中，以研究这种双分子靶向策略的机制。为了证明体内靶向转导的能力，将Jurkat.CD3细胞皮下异种移植到每只小鼠的右侧腹，在注射后8小时将慢病毒载体皮下注射到每只小鼠的两侧。随后，使用无创生物发光成像系统监测报告基因（萤火虫荧光素酶）的表达。
通过在单个慢病毒表面共展示OKT3和融合素，我们可以在体外和体内实现基因向CD3阳性T细胞的靶向递送。
这些结果表明这种工程化慢病毒系统作为细胞类型定向基因递送新工具的潜在用途。
