Corbeau P, Kraus G, Wong-Staal F
Department of Medicine and Biology, University of California at San Diego, La Jolla 92093-0665, USA.
Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):14070-5. doi: 10.1073/pnas.93.24.14070.
By transfecting fibroblast cells with an HIV-1-MN molecular clone with a deletion of the major packaging sequence, we have developed a stable HIV-1 packaging cell line, psi 422, psi 422 cells form syncytia with CD4-positive cells, correctly express HIV-1 structural proteins, and produce a large amount of mature particles with normal reverse transcriptase activity. Yet these particles, in which RNA was not detected by reverse transcriptase-PCR, are not infectious. When stably transfected with an HIV-1-based retroviral vector, the psi 422 cell line produces virions capable of transducing CD4-positive cells with high efficiency (up to 10(5) cells per ml). The availability of this stable noninfectious HIV-1 packaging cell line capable of generating high-titer HIV-1 vectors represents a new step toward the use of an HIV-1 delivery system in gene therapy.
通过用缺失主要包装序列的HIV-1-MN分子克隆转染成纤维细胞，我们构建了一个稳定的HIV-1包装细胞系psi 422。psi 422细胞与CD4阳性细胞形成多核巨细胞，正确表达HIV-1结构蛋白，并产生大量具有正常逆转录酶活性的成熟颗粒。然而，这些颗粒通过逆转录酶-PCR未检测到RNA，不具有感染性。当用基于HIV-1的逆转录病毒载体稳定转染时，psi 422细胞系产生能够高效转导CD4阳性细胞的病毒粒子（每毫升高达10^5个细胞）。这种能够产生高滴度HIV-1载体的稳定的非感染性HIV-1包装细胞系的获得，代表了在基因治疗中使用HIV-1递送系统方面迈出的新一步。
