The Brain Tumor Center, The University of Chicago, Chicago, Illinois, United States of America.
PLoS One. 2011 Mar 28;6(3):e18091. doi: 10.1371/journal.pone.0018091.
Adenoviruses are often used as vehicles to mediate gene delivery for therapeutic purposes, but their research scope in hematological cells remains limited due to a narrow choice of host cells that express the adenoviral receptor (CAR). T cells, which are attractive targets for gene therapy of numerous diseases, remain resistant to adenoviral infection because of the absence of CAR expression. Here, we demonstrate that this resistance can be overcome when murine or human T cells are transduced with an adenovirus incorporating the RGD-fiber modification (Ad-RGD).
METHODOLOGY/PRINCIPAL FINDING: A luciferase-expressing replication-deficient Ad-RGD infected 3-fold higher number of activated primary T cells than an adenovirus lacking the RGD-fiber modification in vitro. Infection with replication-competent Ad-RGD virus also caused increased cell cycling, higher E1A copy number and enriched hexon antigen expression in both human and murine T cells. Transduction with oncolytic Ad-RGD also resulted in higher titers of progeny virus and enhanced the killing of T cells. In vivo, 35-45% of splenic T cells were transduced by Ad-RGD.
Collectively, our results prove that a fiber modified Ad-RGD successfully transduces and replicates in primary T cells of both murine and human origin.
腺病毒常被用作介导基因治疗的载体,但由于能表达腺病毒受体 (CAR) 的宿主细胞选择范围较窄,其在血液细胞中的研究范围有限。由于缺乏 CAR 表达,T 细胞成为多种疾病基因治疗的有吸引力的靶标,但对腺病毒感染仍具有抗性。在这里,我们证明当用包含 RGD 纤维修饰的腺病毒(Ad-RGD)转导鼠或人 T 细胞时,这种抗性可以被克服。
方法/主要发现:体外实验表明,表达荧光素酶的复制缺陷型 Ad-RGD 感染激活的原代 T 细胞的数量比缺乏 RGD 纤维修饰的腺病毒高 3 倍。复制型 Ad-RGD 病毒的感染也导致人源和鼠源 T 细胞中的细胞周期增加、E1A 拷贝数增加和六邻体抗原表达丰富。用溶瘤 Ad-RGD 转导也导致更高滴度的子代病毒,并增强了对 T 细胞的杀伤作用。在体内,Ad-RGD 转导了 35-45%的脾 T 细胞。
总之,我们的研究结果证明纤维修饰的 Ad-RGD 可以成功地转导和复制鼠源和人源的原代 T 细胞。