Valsesia-Wittmann S, Morling F J, Nilson B H, Takeuchi Y, Russell S J, Cosset F L
Centre de Génétique Moléculaire et Cellulaire, Université Claude Bernard Lyon-I, United Kingdom.
J Virol. 1996 Mar;70(3):2059-64. doi: 10.1128/JVI.70.3.2059-2064.1996.
We previously reported a strategy to redirect the retroviral host range by expressing single-chain antibodies (S. J. Russell, R. E. Hawkins, and G. Winter, Nucleic Acids Res. 21:1081-1085, 1993) or ligands (F.-L. Cosset, F. J Morling, Y. Takeuchi, R. A. Weiss, M. K. L. Collins, and S. J. Russell, J. Virol. 69:6314-6322, 1995) at the N terminus of Moloney murine leukemia virus (MoMLV) surface proteins (SU). Although such chimeric envelopes were able to bind the new receptors, the transduction efficiency of retargeted viruses was generally low. We hypothesized that conformational rearrangements of envelope glycoproteins were not optimally triggered following binding, and to overcome these postbinding blocks, we have generated here a set of chimeric MoMLV-derived envelopes targeted to the Ram-1 phosphate transporter in which we have varied the spacing between the Ram-1-binding domain and the MoMLV SU. All of the recombinant envelopes were correctly expressed on virions, and all bound efficiently to Ram-1. However, the interdomain spacing greatly affected the efficiency of gene transfer by retroviral vectors that had bound to Ram-1 via their chimeric envelopes. Optimal interdomain spacing allowed a 100-fold-increased viral transduction via Ram-1 compared to our previous results.
我们之前报道过一种通过在莫洛尼鼠白血病病毒（MoMLV）表面蛋白（SU）的N端表达单链抗体（S. J. 拉塞尔、R. E. 霍金斯和G. 温特，《核酸研究》21:1081 - 1085，1993）或配体（F.-L. 科塞、F. J. 莫林、竹内洋、R. A. 韦斯、M. K. L. 柯林斯和S. J. 拉塞尔，《病毒学杂志》69:6314 - 6322，1995）来改变逆转录病毒宿主范围的策略。尽管这种嵌合包膜能够结合新的受体，但重靶向病毒的转导效率通常较低。我们推测包膜糖蛋白的构象重排在结合后未被最佳触发，为了克服这些结合后障碍，我们在此构建了一组靶向Ram - 1磷酸转运体的源自MoMLV的嵌合包膜，其中我们改变了Ram - 1结合结构域与MoMLV SU之间的间距。所有重组包膜都在病毒粒子上正确表达，并且都能有效结合Ram - 1。然而，结构域间间距极大地影响了通过其嵌合包膜与Ram - 1结合的逆转录病毒载体的基因转移效率。与我们之前的结果相比，最佳的结构域间间距使通过Ram - 1的病毒转导增加了100倍。
