Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
J Virol. 2014 Mar;88(6):3496-504. doi: 10.1128/JVI.02961-13. Epub 2014 Jan 8.
Most human T-lymphotropic virus type 1 (HTLV-1)-infected HeLa and SupT1 cells cease proliferation and become senescent immediately after infection by HTLV-1 or transduction of the HTLV-1 tax gene. The cellular senescence response triggered by Tax is caused by hyperactivated NF-κB and mediated by cyclin-dependent kinase inhibitors, p21(CIP1/WAF1) and p27(KIP1). When NF-κB activity is blocked by a degradation-resistant form of IκBα, ΔN-IκBα, Tax-induced senescence is averted. Here, we show that NF-κB inhibition through the expression of ΔN-IκBα allows cells of a human osteosarcoma (HOS) cell line to be chronically infected by HTLV-1. Stable HTLV-1-producing HOS cell clones can be readily established and isolated. These clones continue to proliferate in culture; express Tax, Rex, Gag, and Env proteins persistently; and transmit HTLV-1 to naive HOS, SupT1, and Jurkat T reporter cell lines readily after cocultivation. As HOS cells are adherent to culture plates, infected T cells in suspension can be easily collected and characterized. The ease with which chronic and productive HTLV-1 infection can be established in cell culture through inhibition of NF-κB affords a useful means to examine in depth the molecular events of HTLV-1 replication and the mechanisms of action of viral genes.
This paper describes a system for establishing cell lines that can be productively infected by human T-lymphotropic virus type 1 (HTLV-1) and can spread HTLV-1 to susceptible cells. Such a system can facilitate the study of HTLV-1 replication in cell culture.
大多数人类 T 淋巴细胞嗜病毒 1（HTLV-1）感染的 HeLa 和 SupT1 细胞在被 HTLV-1 感染或转导 HTLV-1 tax 基因后立即停止增殖并衰老。Tax 引发的细胞衰老反应是由 NF-κB 过度激活引起的，并由细胞周期蛋白依赖性激酶抑制剂 p21（CIP1/WAF1）和 p27（KIP1）介导。当 NF-κB 活性被降解抗性形式的 IκBα，ΔN-IκBα 阻断时，Tax 诱导的衰老就会被避免。在这里，我们表明通过表达 ΔN-IκBα 抑制 NF-κB 可以使人类骨肉瘤（HOS）细胞系的细胞被 HTLV-1 慢性感染。可以轻易建立和分离稳定产生 HTLV-1 的 HOS 细胞克隆。这些克隆在培养物中继续增殖；持续表达 Tax、Rex、Gag 和 Env 蛋白；并在共培养后容易将 HTLV-1 传递给幼稚的 HOS、SupT1 和 Jurkat T 报告细胞系。由于 HOS 细胞附着在培养板上，悬浮感染的 T 细胞可以很容易地收集和鉴定。通过抑制 NF-κB 轻易地在细胞培养中建立慢性和有生产力的 HTLV-1 感染，为深入研究 HTLV-1 复制的分子事件和病毒基因的作用机制提供了一种有用的手段。
本文描述了一种建立可被人类 T 淋巴细胞嗜病毒 1（HTLV-1）有效感染并可将 HTLV-1 传播给易感细胞的细胞系的系统。这样的系统可以促进在细胞培养中研究 HTLV-1 复制。
