Laboratory of Biology and Modeling of the Cell, Ecole Normale Supérieure (ENS) de Lyon, INSERM U1210, CNRS UMR5239, 46 allée d'Italie, Lyon, France.
International Center for Infectiology Research, ENS de Lyon, Université Claude Bernard Lyon 1, INSERM U1111, CNRS UMR 5308, 46 allée d'Italie, Lyon, France.
PLoS Pathog. 2018 Mar 22;14(3):e1006933. doi: 10.1371/journal.ppat.1006933. eCollection 2018 Mar.
Human T-cell leukemia virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL), an aggressive malignant proliferation of activated CD4+ T lymphocytes. The viral Tax oncoprotein is critically involved in both HTLV-1-replication and T-cell proliferation, a prerequisite to the development of ATLL. In this study, we investigated the in vivo contribution of the Tax PDZ domain-binding motif (PBM) to the lymphoproliferative process. To that aim, we examined T-cell proliferation in humanized mice (hu-mice) carrying a human hemato-lymphoid system infected with either a wild type (WT) or a Tax PBM-deleted (ΔPBM) provirus. We observed that the frequency of CD4+ activated T-cells in the peripheral blood and in the spleen was significantly higher in WT than in ΔPBM hu-mice. Likewise, human T-cells collected from WT hu-mice and cultivated in vitro in presence of interleukin-2 were proliferating at a higher level than those from ΔPBM animals. We next examined the association of Tax with the Scribble PDZ protein, a prominent regulator of T-cell polarity, in human T-cells analyzed either after ex vivo isolation or after in vitro culture. We confirmed the interaction of Tax with Scribble only in T-cells from the WT hu-mice. This association correlated with the presence of both proteins in aggregates at the leading edge of the cells and with the formation of long actin filopods. Finally, data from a comparative genome-wide transcriptomic analysis suggested that the PBM-PDZ association is implicated in the expression of genes regulating proliferation, apoptosis and cytoskeletal organization. Collectively, our findings suggest that the Tax PBM is an auxiliary motif that contributes to the sustained growth of HTLV-1 infected T-cells in vivo and in vitro and is essential to T-cell immortalization.
人类 T 细胞白血病病毒 1 型(HTLV-1)是成人 T 细胞白血病/淋巴瘤(ATLL)的病原体,是一种激活的 CD4+T 淋巴细胞的侵袭性恶性增殖。病毒 Tax 癌蛋白在 HTLV-1 复制和 T 细胞增殖中都起着至关重要的作用,这是 ATLL 发展的前提。在这项研究中,我们研究了 Tax PDZ 结构域结合基序(PBM)在淋巴增殖过程中的体内贡献。为此,我们检查了携带受感染的人类血液 - 淋巴系统的人源化小鼠(hu-mice)中的 T 细胞增殖,该系统感染了野生型(WT)或 Tax PBM 缺失(ΔPBM)前病毒。我们观察到,外周血和脾脏中 CD4+激活 T 细胞的频率在 WT 比在 ΔPBM hu-mice 中明显更高。同样,从 WT hu-mice 中收集并在存在白细胞介素-2 的情况下在体外培养的人 T 细胞比从 ΔPBM 动物中收集的 T 细胞增殖水平更高。接下来,我们检查了 Tax 与 Scribble PDZ 蛋白的关联,Scribble PDZ 蛋白是 T 细胞极性的主要调节剂,我们在分析体外分离或体外培养后的人 T 细胞时,都检查了 Tax 与 Scribble 的关联。我们仅在 WT hu-mice 的 T 细胞中证实了 Tax 与 Scribble 的相互作用。这种关联与两种蛋白在细胞前缘的聚集体中以及长肌动蛋白丝状伪足的形成相关。最后,比较基因组范围的转录组分析数据表明,PBM-PDZ 关联参与了调节增殖、凋亡和细胞骨架组织的基因表达。总的来说,我们的发现表明,Tax PBM 是一种辅助基序,有助于 HTLV-1 感染的 T 细胞在体内和体外的持续生长,并对 T 细胞永生化至关重要。