Protection of herpes simplex virus thymidine kinase-transduced cells from ganciclovir-mediated cytotoxicity by bystander cells: the Good Samaritan effect.
Protection of herpes simplex virus thymidine kinase-transduced cells from ganciclovir-mediated cytotoxicity by bystander cells: the Good Samaritan effect.
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发表时间：
1997-05
期刊：
影响因子：
11.2
通讯作者：
M. Wygoda;Melinda R. Wilson;Mary A Davis;J. Trosko;A. Rehemtulla;T. Lawrence
中科院分区：
文献类型：
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作者：
M. Wygoda;Melinda R. Wilson;Mary A Davis;J. Trosko;A. Rehemtulla;T. Lawrence
虽然基因治疗领域的相当多的注意力都集中在阐明转导细胞可以杀死旁观者细胞的机制上，但对于旁观者细胞如何影响转导细胞却知之甚少。我们假设旁观者细胞，特别是如果它们能够进行缝隙连接通讯，可以保护转导单纯疱疹病毒胸苷激酶(HSV-TK)的细胞免受更昔洛韦(GCV)诱导的细胞毒性。为了验证这一假设，我们使用了能够进行有效缝隙连接通讯的大鼠肝细胞系(WB)、转导HSV-TK的WB克隆(WB-TK)和通讯功能不全的WB细胞亚克隆(AB1)。我们将WB-TK细胞与WB或AB1细胞共培养，用GCV处理它们，然后将这些细胞接种到选择性培养液中，使我们能够独立地量化WB-TK细胞或旁观者细胞的存活比例。我们发现，WB旁观者细胞对GCV处理的WB-TK细胞具有1000倍的保护作用。AB1细胞提供了可检测到的保护，但明显较少。这些发现表明，单纯疱疹病毒胸苷激酶转导的细胞可以显著地受到旁观者细胞的保护，特别是那些能够进行缝隙连接通讯的细胞。这种“好心人”效应是通过延长有毒代谢物来源的存活来提高基因治疗的整体疗效，还是通过提高转导细胞的存活来降低疗效，还需要在体内确定。
Although considerable attention has been directed in the field of gene therapy toward elucidating the mechanism by which a transduced cell could kill a bystander cell, little is known about how bystander cells may affect transduced cells. We hypothesized that bystander cells, particularly if they were capable of gap junctional communication, could protect cells transduced with the herpes simplex virus thymidine kinase (HSV-TK) from ganciclovir (GCV)-induced cytotoxicity. To test this hypothesis, we used a rat hepatocyte cell line (WB) that can carry out efficient gap junctional communication, a WB clone transduced with HSV-TK (WB-TK), and a communication-incompetent subclone of WB cells (aB1). We cocultured WB-TK cells with either WB or aB1 cells, treated them with GCV, and then plated the cells into selective media that permitted us to quantify independently the surviving fraction of WB-TK cells or bystander cells. We found that WB bystander cells conferred up to a 1000-fold protection on WB-TK cells treated with GCV. aB1 cells conferred detectable, but significantly less, protection. These findings demonstrate that herpes simplex virus thymidine kinase-transduced cells can be significantly protected by bystander cells, particularly those that can carry out gap junctional communication. Whether this "Good Samaritan" effect improves the overall efficacy of gene therapy, by prolonging the survival of the source of toxic metabolites, or decreases effectiveness by increasing the survival of transduced cells will need to be determined in vivo.