Herpes simplex virus targeting to the EGF receptor by a gD-specific soluble bridging molecule
Herpes simplex virus targeting to the EGF receptor by a gD-specific soluble bridging molecule
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DOI：
10.1016/j.ymthe.2004.12.012
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发表时间：
2005-04-01
影响因子：
12.4
通讯作者：
Glorioso, JC
中科院分区：
文献类型：
--
作者：
Nakano, K;Asano, R;Glorioso, JC
单纯疱疹病毒1型（HSV-1）通过包膜糖蛋白（g）C和B与细胞表面糖胺聚糖（GAG）的初始结合以及随后涉及包膜913、g B和gH/gL的膜融合进入细胞。目前的见解表明，融合过程是由gD与同源细胞受体（如广泛分布的细胞粘附分子nectin-1）相互作用引发的。为了重定向HSV-1的向性，我们已经产生了可溶性衔接蛋白（P-V528 LH），其包含与识别EGF受体的单链抗体（528 LH）融合的连接蛋白-1的gD结合可变结构域。衔接子分子使HSV-1能够进入表达人EGF受体的天然非允许性CHO细胞，但不能进入缺乏受体的CHO细胞，并且当衔接子的抗体部分被不同特异性的抗体替换时，未观察到进入。衔接子介导的进入随着病毒剂量的增加而增加，并且几乎与病毒直接进入带有连接蛋白-1的CHO细胞一样有效。进入依赖于病毒gD，并在没有细胞GAG的情况下减少。这些实验代表了可溶性分子可以通过新受体指导HSV感染的第一个证明，支持这种方法用于HSV重靶向的可能效用。
Herpes simplex virus 1 (HSV-1) enters cells via initial binding of envelope glycoproteins (g) C and B to cell-surface glycosaminoglycans (GAGs) and subsequent membrane fusion involving envelope 913, gB, and gH/gL. Current insights suggest that the fusion process is initiated by interaction of gD with a cognate cellular receptor, such as the widely distributed cell adhesion molecule nectin-1. To redirect the tropism of HSV-1, we have generated a soluble adapter protein (P-V528LH) comprising the gD-binding variable domain of nectin-1 fused to a single-chain antibody (528LH) recognizing the EGF receptor. The adapter molecule enabled HSV-1 entry into naturally nonpermissive CHO cells expressing the human EGF receptor, but not into CHO cells lacking the receptor, and entry was not observed when the antibody portion of the adapter was replaced with an antibody of different specificity. Adapter-mediated entry increased with the viral dose and was nearly as efficient as direct viral entry into nectin-1-bearing CHO cells. Entry depended on viral gD and was diminished in the absence of cellular GAGs. These experiments represent the first demonstration that a soluble molecule can direct HSV infection via a new receptor, supporting the possible utility of this approach for HSV retargeting.