Cytokine signals are sufficient for HIV-1 infection of resting human T lymphocytes.
Cytokine signals are sufficient for HIV-1 infection of resting human T lymphocytes.
复制标题
DOI：
10.1084/jem.189.11.1735
复制
发表时间：
1999-06-07
期刊：
影响因子：
--
通讯作者：
Littman DR
中科院分区：
文献类型：
--
作者：
Unutmaz D;KewalRamani VN;Marmon S;Littman DR
慢病毒载体被认为是在未分裂的原代细胞中传递和稳定表达基因的有效载体。然而，某些细胞类型，如静息T淋巴细胞，对感染HIV-1具有抵抗力。建立稳定的基因输送到原代人类淋巴细胞的参数和克服静止T细胞对HIV感染的抵抗力的方法，可能会使潜在的基因治疗应用、体外原代细胞的基因研究以及更好地了解慢病毒生命周期的各个阶段。在这里，我们证明了HIV-1衍生的载体可以用于将基因稳定地输送到激活的人类T细胞以及自然杀伤细胞和树突状细胞中。值得注意的是，在没有任何其他刺激的情况下，当与细胞因子IL(IL)-2、IL-4、IL-7或IL-15培养时，相当大一部分静止的T细胞可以稳定地与HIV-1载体一起转导，或者在较低水平的IL-6的作用下，可以稳定地转导静息T细胞。用这些细胞因子刺激的静息T细胞也可以感染复制能力强的HIV-1。为了测试该系统在原代T细胞中进行结构功能分析的有效性，我们将野生型和突变型小鼠CD28导入人T细胞，并显示了共刺激信号中对CD28胞浆结构域的要求。在原代T细胞中稳定表达感兴趣基因的能力将是一种有价值的工具，用于遗传和结构功能研究，而这些研究以前仅限于转化的细胞系。此外，细胞因子信号足以允许静息T细胞与HIV的转导，这一发现可能与理解HIV-1的传播机制和发病机制有关。
Lentiviral vectors have been advocated to be effective vehicles for the delivery and stable expression of genes in nondividing primary cells. However, certain cell types, such as resting T lymphocytes, are resistant to infection with HIV-1. Establishing parameters for stable gene delivery into primary human lymphocytes and approaches to overcome the resistance of resting T cells to HIV infection may permit potential gene therapy applications, genetic studies of primary cells in vitro, and a better understanding of the stages of the lentiviral life cycle. Here we demonstrate that an HIV-1–derived vector can be used for stable delivery of genes into activated human T cells as well as natural killer and dendritic cells. Remarkably, a sizeable fraction of resting T cells was stably transduced with the HIV-1 vector when cultured with the cytokine interleukin (IL)-2, IL-4, IL-7, or IL-15, or, at a lower level, with IL-6, in the absence of any other stimuli. Resting T cells stimulated with these cytokines could also be infected with replication-competent HIV-1. To test the utility of this system for performing structure–function analysis in primary T cells, we introduced wild-type as well as a mutant form of murine CD28 into human T cells and showed a requirement for the CD28 cytoplasmic domain in costimulatory signaling. The ability to stably express genes of interest in primary T cells will be a valuable tool for genetic and structure–function studies that previously have been limited to transformed cell lines. In addition, the finding that cytokine signals are sufficient to permit transduction of resting T cells with HIV may be relevant for understanding mechanism of HIV-1 transmission and pathogenesis.