OBJECTIVES:To evaluate in vitro and in vivo a strategy for gene therapy for AIDS based on the transfer on interferon (IFN)-alpha, -beta and -gamma genes to human cells.

DESIGN:Human U937 promonocytic cells were stably transfected with Tat-inducible IFN expression vectors conferring an antiviral state against infection with HIV.

METHODS:Transfected cells were either infected by HIV-1 in vitro or transplanted into severe combined immunodeficient (SCID) mice for an HIV challenge in vivo.

RESULTS:U937 cell lines stably carrying IFN transgenes under the positive control of the HIV-1 Tat protein were highly resistant to HIV-1 replication in vitro. This antiviral resistance was associated with a strong induction of IFN synthesis immediately following the viral infection. HIV-1 proteins were found to be specifically trapped within the genetically modified cells. In contrast, all IFN-U937 cells permitted full HIV-2 replication. Transfected cells injected into SCID mice and challenged against HIV-1 were strongly resistant to infection when cells were transduced with IFN-alpha of IFN-beta genes. However, IFN-gamma-transfected cells permitted HIV-1 infection in vivo despite the induction of a high level of IFN-gamma secretion. The quantity of proviral DNA was 10(5)-fold lower in IFN-alpha- or IFN-beta-transfected U937 cells collected from these SCID mice than that in non-transfected cells.

CONCLUSIONS:Our results substantiated the validity of a strategy, bases on the transfer of HIV-1-inducible IFN-alpha or IFN-beta genes, to confer antiviral resistance to human cells.

译文

目标 : 基于干扰素 (IFN)-α,-β 和-γ 基因向人类细胞的转移,在体外和体内评估用于艾滋病的基因治疗策略。
设计 : 人U937单核细胞用Tat诱导的IFN表达载体稳定转染,赋予抗HIV感染的抗病毒状态。
方法 : 转染的细胞要么通过体外HIV-1感染,要么移植到严重的联合免疫缺陷 (SCID) 小鼠体内进行HIV攻击。
结果 : 在HIV-1 Tat蛋白的阳性控制下稳定携带IFN转基因的U937细胞系在体外对HIV-1复制具有高度抗性。这种抗病毒耐药性与病毒感染后立即强烈诱导IFN合成有关。发现HIV-1蛋白质被特异性地捕获在转基因细胞内。相反,所有IFN-U937细胞允许完全HIV-2复制。当用IFN-β 基因的IFN-α 转导细胞时,注入SCID小鼠并对HIV-1进行攻击的转染细胞对感染具有强烈抗性。然而,尽管诱导了高水平的IFN-γ 分泌,但转染IFN-γ 的细胞允许体内HIV-1感染。从这些SCID小鼠收集的IFN-α 或IFN-β 转染的U937细胞中前病毒DNA的数量比未转染的细胞低10(5) 倍。
结论 : 我们的结果证实了基于HIV-1-inducible IFN-α 或IFN-β 基因转移的策略对人类细胞具有抗病毒抗性的有效性。

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