Strang Blair L, Takeuchi Yasuhiro, Relander Thomas, Richter Johan, Bailey Ranbir, Sanders David A, Collins Mary K L, Ikeda Yasuhiro
Department of Immunology and Molecular Pathology, University College London, University of London, London, United Kingdom.
J Virol. 2005 Feb;79(3):1765-71. doi: 10.1128/JVI.79.3.1765-1771.2005.
Alphavirus glycoproteins have broad host ranges. Human immunodeficiency virus type 1 (HIV-1) vectors pseudotyped with their glycoproteins could extend the range of tissues that can be transduced in both humans and animal models. Here, we established stable producer cell lines for HIV vectors pseudotyped with alphavirus Ross River virus (RRV) and Semliki Forest virus (SFV) glycoproteins E2E1. RRV E2E1-stable clones could routinely produce high-titer pseudotyped vectors for at least 5 months. SFV E2E1-stable clones, however, produced relatively low titers. We examined the properties of RRV E2E1-pseudotyped vectors [HIV-1(RRV)] and compared them with amphotropic murine leukemia virus Env- and vesicular stomatitis virus glycoprotein G-pseudotyped vectors. HIV-1(RRV) displayed a number of characteristics which would be advantageous in ex vivo and in vivo experiments, including resistance to inactivation by heat-labile components in fresh human sera and thermostability at 37 degrees C. Upon single-step concentration by ultracentrifugation of HIV-1(RRV), we could achieve vector stocks with titers up to 6 x 10(7) IU/ml. HIV-1(RRV) efficiently transduced cells from several different species, including murine primary dendritic cells, but failed to transduce human and murine T cells as well as human hematopoietic stem cells (HSC). These results indicate that HIV-1(RRV) could be used in a number of applications including animal model experiments and suggest that expression of RRV cellular receptors is limited or absent in certain cell types such as T cells and human HSC.
甲病毒糖蛋白具有广泛的宿主范围。用其糖蛋白假型化的1型人类免疫缺陷病毒（HIV-1）载体可扩大在人类和动物模型中能够被转导的组织范围。在此，我们建立了用甲病毒罗斯河病毒（RRV）和塞姆利基森林病毒（SFV）糖蛋白E2E1假型化的HIV载体的稳定生产细胞系。RRV E2E1稳定克隆可常规产生高滴度假型化载体至少5个月。然而，SFV E2E1稳定克隆产生的滴度相对较低。我们检测了RRV E2E1假型化载体[HIV-1（RRV）]的特性，并将其与嗜双性鼠白血病病毒Env和水疱性口炎病毒糖蛋白G假型化载体进行比较。HIV-1（RRV）表现出一些在体外和体内实验中具有优势的特性，包括对新鲜人血清中热不稳定成分的灭活具有抗性以及在37℃下的热稳定性。通过对HIV-1（RRV）进行超速离心单步浓缩，我们可获得滴度高达6×10⁷ IU/ml的载体储备液。HIV-1（RRV）能有效转导来自几种不同物种的细胞，包括小鼠原代树突状细胞，但未能转导人和小鼠T细胞以及人造血干细胞（HSC）。这些结果表明，HIV-1（RRV）可用于包括动物模型实验在内的多种应用，并提示RRV细胞受体在某些细胞类型如T细胞和人HSC中表达受限或缺失。
