Department of Microbiology, Immunology & Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
J Virol. 2012 Jan;86(1):373-81. doi: 10.1128/JVI.05411-11. Epub 2011 Oct 19.
Here we report a novel viral glycoprotein created by replacing a natural receptor-binding sequence of the ecotropic Moloney murine leukemia virus envelope glycoprotein with the peptide ligand somatostatin. This new chimeric glycoprotein, which has been named the Sst receptor binding site (Sst-RBS), gives targeted transduction based on three criteria: (i) a gain of the use of a new entry receptor not used by any known virus; (ii) targeted entry at levels comparable to gene delivery by wild-type ecotropic Moloney murine leukemia virus and vesicular stomatitis virus (VSV) G glycoproteins; and (iii) a loss of the use of the natural ecotropic virus receptor. Retroviral vectors coated with Sst-RBS gained the ability to bind and transduce human 293 cells expressing somatostatin receptors. Their infection was specific to target somatostatin receptors, since a synthetic somatostatin peptide inhibited infection in a dose-dependent manner and the ability to transduce mouse cells bearing the natural ecotropic receptor was effectively lost. Importantly, vectors coated with the Sst-RBS glycoprotein gave targeted entry of up to 1 × 10(6) transducing U/ml, a level comparable to that seen with infection of vectors coated with the parental wild-type ecotropic Moloney murine leukemia virus glycoprotein through the ecotropic receptor and approaching that of infection of VSV G-coated vectors through the VSV receptor. To our knowledge, this is the first example of a glycoprotein that gives targeted entry of retroviral vectors at levels comparable to the natural capacity of viral envelope glycoproteins.
在这里,我们报告了一种新型的病毒糖蛋白,它通过用生长抑素肽取代嗜性 Moloney 鼠白血病病毒包膜糖蛋白的天然受体结合序列来创建。这种新的嵌合糖蛋白已被命名为生长抑素受体结合位点(Sst-RBS),基于三个标准进行靶向转导:(i)获得使用任何已知病毒都不使用的新进入受体;(ii)靶向进入水平与野生型嗜性 Moloney 鼠白血病病毒和水疱性口炎病毒(VSV)G 糖蛋白的基因递送相当;(iii)丧失使用天然嗜性病毒受体的能力。用 Sst-RBS 包被的逆转录病毒载体获得了与表达生长抑素受体的人 293 细胞结合和转导的能力。它们的感染是特异性针对靶生长抑素受体的,因为合成的生长抑素肽以剂量依赖的方式抑制感染,并且有效丧失了转导携带天然嗜性受体的小鼠细胞的能力。重要的是,用 Sst-RBS 糖蛋白包被的载体可进行高达 1×106 转导 U/ml 的靶向进入,这一水平与通过嗜性受体感染用亲本野生型嗜性 Moloney 鼠白血病病毒糖蛋白包被的载体的感染相当,接近通过 VSV 受体感染用 VSV G 包被的载体的感染。据我们所知,这是第一个能够使逆转录病毒载体以与天然包膜糖蛋白的天然能力相当的水平进行靶向进入的糖蛋白的例子。