Miyake K, Suzuki N, Matsuoka H, Tohyama T, Shimada T
Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo, Japan.
Hum Gene Ther. 1998 Mar 1;9(4):467-75. doi: 10.1089/hum.1998.9.4-467.
Human immunodeficiency virus type 1 (HIV-1)-based vectors are thought to be useful for gene transfer into nondividing cells. We examined whether HIV vectors can really integrate into the chromosomes of nondividing cells. CD4+HeLa cells arrested at the G2 or G1/S phase were incubated with the HIV vector pseudotyped with the HIV envelope. The transduction efficiency of the HIV vector in these nondividing cells was comparable to that in proliferating cells. Sequencing of the polymerase chain reaction-amplified fragments containing the junction sites showed that the HIV vector was stably integrated into the chromosomal DNA. It was also demonstrated that terminally differentiated human macrophages and nonproliferating NT neurons could be transduced by the HIV vector after adenovirus-mediated expression of CD4. These results suggest that the HIV vector may be useful not only for gene therapy of AIDS but also for a variety of gene therapy protocols targeting nondividing cells.
基于1型人类免疫缺陷病毒（HIV-1）的载体被认为可用于将基因导入非分裂细胞。我们研究了HIV载体是否真的能整合到非分裂细胞的染色体中。将停滞在G2或G1/S期的CD4⁺HeLa细胞与用HIV包膜假型化的HIV载体一起孵育。HIV载体在这些非分裂细胞中的转导效率与在增殖细胞中的转导效率相当。对包含连接位点的聚合酶链反应扩增片段进行测序表明，HIV载体稳定地整合到了染色体DNA中。还证明，在腺病毒介导的CD4表达后，终末分化的人类巨噬细胞和非增殖性NT神经元可被HIV载体转导。这些结果表明，HIV载体不仅可能对艾滋病的基因治疗有用，而且对针对非分裂细胞的各种基因治疗方案也可能有用。
