Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
PLoS Pathog. 2011 Jun;7(6):e1002088. doi: 10.1371/journal.ppat.1002088. Epub 2011 Jun 16.
Human parvovirus B19 (B19V) causes a variety of human diseases. Disease outcomes of bone marrow failure in patients with high turnover of red blood cells and immunocompromised conditions, and fetal hydrops in pregnant women are resulted from the targeting and destruction of specifically erythroid progenitors of the human bone marrow by B19V. Although the ex vivo expanded erythroid progenitor cells recently used for studies of B19V infection are highly permissive, they produce progeny viruses inefficiently. In the current study, we aimed to identify the mechanism that underlies productive B19V infection of erythroid progenitor cells cultured in a physiologically relevant environment. Here, we demonstrate an effective reverse genetic system of B19V, and that B19V infection of ex vivo expanded erythroid progenitor cells at 1% O(2) (hypoxia) produces progeny viruses continuously and efficiently at a level of approximately 10 times higher than that seen in the context of normoxia. With regard to mechanism, we show that hypoxia promotes replication of the B19V genome within the nucleus, and that this is independent of the canonical PHD/HIFα pathway, but dependent on STAT5A and MEK/ERK signaling. We further show that simultaneous upregulation of STAT5A signaling and down-regulation of MEK/ERK signaling boosts the level of B19V infection in erythroid progenitor cells under normoxia to that in cells under hypoxia. We conclude that B19V infection of ex vivo expanded erythroid progenitor cells at hypoxia closely mimics native infection of erythroid progenitors in human bone marrow, maintains erythroid progenitors at a stage conducive to efficient production of progeny viruses, and is regulated by the STAT5A and MEK/ERK pathways.
人细小病毒 B19(B19V)可引起多种人类疾病。在红血球周转率高和免疫功能低下的患者中骨髓衰竭以及孕妇胎儿水肿的疾病结果,是由于 B19V 靶向和破坏人类骨髓中特定的红系祖细胞所致。尽管最近用于 B19V 感染研究的体外扩增的红系祖细胞具有高度的易感性,但它们产生的子代病毒效率低下。在本研究中,我们旨在确定在生理相关环境中培养的红系祖细胞中有效产生 B19V 感染的机制。在这里,我们展示了 B19V 的有效反向遗传系统,并且在 1% O(2)(缺氧)下对体外扩增的红系祖细胞进行 B19V 感染可连续有效地产生子代病毒,其效率约比在正常氧条件下高 10 倍。就机制而言,我们表明缺氧促进 B19V 基因组在核内的复制,并且这独立于经典的 PHD/HIFα途径,而是依赖于 STAT5A 和 MEK/ERK 信号。我们进一步表明,同时上调 STAT5A 信号和下调 MEK/ERK 信号可将红系祖细胞在正常氧条件下的 B19V 感染水平提高到缺氧条件下的水平。我们得出结论,在低氧条件下对体外扩增的红系祖细胞进行 B19V 感染,可紧密模拟人类骨髓中红系祖细胞的天然感染,使红系祖细胞保持在有利于高效产生子代病毒的阶段,并受 STAT5A 和 MEK/ERK 途径调节。