Yang G, Zhong Q, Huang W, Reiser J, Schwarzenberger P
Department of Genetics, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA.
Cancer Gene Ther. 2006 May;13(5):460-8. doi: 10.1038/sj.cgt.7700911.
In principle, transient nongenetic modification of a noninfectious gene transfer virus enabling a one time infection and transduction of human cells could eliminate the risk of formation of replication competent virus. Formation of a molecular conjugate vector by conjugation of noninfective ecotropic murine Moloney leukemia virus to polylysine (eMMLV-PL) enabled high-efficiency transduction of human HPC using in vitro and in vivo assays. Xenotransplanted NOD-SCID mice durably expressed the transgene in human leukocytes and human progenitor cells with eMMLV-PL achieving three-fold increased transduction efficiency when directly compared to optimized amphotropic MMLV (aMMLV) transduction. Both aMMLV and eMMLV assembled conjugate vectors showed similar transduction efficiency indicating predominant polylysine-mediated uptake. Integration of retroviral sequences was determined from individual human HPC recovered from eMMLV-PL-xenotransplanted animals. This simple and versatile concept of conjugate gene transfer vectors has the potential to enhance transduction efficiency as well as to improve certain safety aspects of human gene therapy. Moreover, because it permits effective cellular internalization of particles, this concept of molecular conjugates can be used as research tool to investigate the interactions of otherwise noninfectious viruses or modified viral particles at the genomic level.
原则上，对非感染性基因转移病毒进行短暂的非基因改造，使其能够一次性感染和转导人类细胞，可消除产生具有复制能力病毒的风险。通过将非感染性嗜亲性莫洛尼氏鼠白血病病毒与聚赖氨酸结合形成分子共轭载体（eMMLV-PL），利用体外和体内试验实现了对人造血祖细胞（HPC）的高效转导。异种移植的NOD-SCID小鼠在人类白细胞和人类祖细胞中持久表达转基因，与优化后的双嗜性莫洛尼氏鼠白血病病毒（aMMLV）转导相比，eMMLV-PL的转导效率提高了三倍。aMMLV和eMMLV组装的共轭载体显示出相似的转导效率，表明主要是聚赖氨酸介导的摄取。从eMMLV-PL异种移植动物中回收的单个人类HPC中确定了逆转录病毒序列的整合情况。这种简单通用的共轭基因转移载体概念有潜力提高转导效率，并改善人类基因治疗的某些安全方面。此外，由于它允许颗粒有效地内化进入细胞，这种分子共轭概念可作为研究工具，在基因组水平上研究其他非感染性病毒或修饰病毒颗粒的相互作用。
