Weerasinghe M, Liem S E, Asad S, Read S E, Joshi S
Department of Microbiology, University of Toronto, Ontario, Canada.
J Virol. 1991 Oct;65(10):5531-4. doi: 10.1128/JVI.65.10.5531-5534.1991.
Toward gene therapy for the treatment of human immunodeficiency virus type 1 (HIV-1) infections in AIDS, Moloney murine leukemia virus-derived retroviral vectors were engineered to allow constitutive and tat-inducible expression of an HIV-1 5' leader sequence-specific ribozyme (Rz1). These vectors were used to infect the human CD4+ lymphocyte-derived MT4 cell line. The stable MT4 transformants expressing an HIV-1 RNA-specific ribozyme, under the control of the herpes simplex virus thymidine kinase (tk) promoter, were found to be somewhat resistant to HIV-1 infection as virus production was delayed. In cells allowing ribozyme expression under control of the simian virus 40 or cytomegalovirus promoter, the rate of HIV-1 multiplication was slightly decreased, and virus production was delayed by about 14 days. The highest level of resistance to HIV-1 infection was observed in MT4 cells transformed with a vector containing a fusion tk-TAR (trans activation-responsive) promoter to allow ribozyme expression in a constitutive and tat-inducible manner; no HIV-1 production was observed 22 days after infection of these cells. These results indicate that retroviral vectors expressing HIV-1 RNA-specific ribozymes can be used to confer resistance to HIV-1 infection.
为了将基因治疗用于治疗艾滋病中的人类免疫缺陷病毒1型(HIV-1)感染,对莫洛尼鼠白血病病毒衍生的逆转录病毒载体进行了改造,使其能够组成型和tat诱导型表达HIV-1 5'前导序列特异性核酶(Rz1)。这些载体被用于感染人CD4+淋巴细胞衍生的MT4细胞系。发现在单纯疱疹病毒胸苷激酶(tk)启动子控制下表达HIV-1 RNA特异性核酶的稳定MT4转化体对HIV-1感染有一定抗性,因为病毒产生被延迟。在猿猴病毒40或巨细胞病毒启动子控制下允许核酶表达的细胞中,HIV-1增殖速率略有降低,病毒产生延迟约14天。在用含有融合tk-TAR(反式激活应答元件)启动子的载体转化的MT4细胞中观察到对HIV-1感染的最高抗性水平,该启动子允许以组成型和tat诱导型方式表达核酶;在感染这些细胞22天后未观察到HIV-1产生。这些结果表明,表达HIV-1 RNA特异性核酶的逆转录病毒载体可用于赋予对HIV-1感染的抗性。