Rift Valley fever virus 78kDa envelope protein attenuates virus replication in macrophage-derived cell lines and viral virulence in mice.
Rift Valley fever virus 78kDa envelope protein attenuates virus replication in macrophage-derived cell lines and viral virulence in mice.
复制标题
裂谷热病毒78kDa包膜蛋白减弱巨噬细胞来源的细胞系中的病毒复制和小鼠中的病毒毒力
DOI：
10.1371/journal.pntd.0009785
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发表时间：
2021-09
影响因子：
3.8
通讯作者：
Makino S
中科院分区：
文献类型：
--
作者：
Terasaki K;Kalveram B;Johnson KN;Juelich T;Smith JK;Zhang L;Freiberg AN;Makino S
裂谷热病毒（RVFV）是一种由蚊子传播的布尼亚病毒，宿主范围广泛，包括反刍动物和人类。RVFV的爆发对非洲国家的公共卫生和畜牧业产生了破坏性影响。然而，在非流行国家没有批准用于人用的RVFV疫苗，也没有FDA批准用于RVFV治疗的抗病毒药物。RVFV 78 kDa蛋白（P78）是一种膜糖蛋白，在病毒在蚊子宿主中的传播中起作用，但其在哺乳动物宿主中的生物学作用尚不清楚。我们产生了减毒的RVFV MP-12株衍生的P78-High病毒和强毒的ZH 501株衍生的ZH 501-P78-High病毒，与它们的亲本病毒相比，这两种病毒都表达更高水平的P78并且在病毒粒子中携带更高水平的P78。我们还产生了另一种不表达P78的MP-12衍生的突变病毒（P78-KO病毒）。MP-12和P78-KO病毒在成纤维细胞系和Huh 7细胞中复制到相似的水平，而P78-High病毒在Vero E6细胞、成纤维细胞系和Huh 7细胞中的复制优于MP-12。值得注意的是，P78-High病毒和P78-KO病毒在巨噬细胞系中的复制效率分别低于MP-12和更高。ZH 501-P78-高病毒在巨噬细胞系中也复制不良。我们的数据进一步表明，P78-High病毒与细胞的无效结合导致病毒内化无效、病毒感染性低和巨噬细胞系中病毒复制减少。P78-High病毒和P78-KO病毒在幼龄小鼠中的毒力分别低于MP-12和高于MP-12。ZH 501-P78-High病毒在小鼠中的毒力也低于ZH 501。这些数据表明，高水平的P78表达通过阻止病毒在巨噬细胞中的有效复制来减弱RVFV毒力。基因改变导致P78表达增加可能作为一种新的策略，为RVFV毒力的衰减和安全的RVFV疫苗的产生。裂谷热病毒（RVFV）是裂谷热的病原体，主要在非洲国家流行。该病毒感染多种哺乳动物物种，包括反刍动物和人类，并对公共卫生和畜牧业造成破坏性影响。由于RVFV是由几种在非流行地区普遍存在的蚊子传播的，因此该病毒存在传播到目前流行地区以外的潜在风险。然而，没有有效的治疗方法，也没有商业上可获得的疫苗供人类使用。RVFV编码一个78 kDa的蛋白质（P78），在哺乳动物细胞中的生物学功能尚未确定。我们发现，表达更高水平的P78的突变体RVFV（P78-高病毒）在巨噬细胞系中表现出比亲本RVFV更低的感染性和更低的复制效率。P78-高病毒和缺乏P78表达的突变体RVFV在小鼠中分别显示出比亲本病毒更低和更高的毒力，表明高水平的P78表达阻止了病毒在巨噬细胞中的有效复制，导致病毒毒力的减弱。导致P78表达增加的遗传改变可作为产生作为疫苗候选物的减毒RVFV的新策略。
Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus with a wide host range including ruminants and humans. RVFV outbreaks have had devastating effects on public health and the livestock industry in African countries. However, there is no approved RVFV vaccine for human use in non-endemic countries and no FDA-approved antiviral drug for RVFV treatment. The RVFV 78kDa protein (P78), which is a membrane glycoprotein, plays a role in virus dissemination in the mosquito host, but its biological role in mammalian hosts remains unknown. We generated an attenuated RVFV MP-12 strain-derived P78-High virus and a virulent ZH501 strain-derived ZH501-P78-High virus, both of which expressed a higher level of P78 and carried higher levels of P78 in the virion compared to their parental viruses. We also generated another MP-12-derived mutant virus (P78-KO virus) that does not express P78. MP-12 and P78-KO virus replicated to similar levels in fibroblast cell lines and Huh7 cells, while P78-High virus replicated better than MP-12 in Vero E6 cells, fibroblast cell lines, and Huh7 cells. Notably, P78-High virus and P78-KO virus replicated less efficiently and more efficiently, respectively, than MP-12 in macrophage cell lines. ZH501-P78-High virus also replicated poorly in macrophage cell lines. Our data further suggest that inefficient binding of P78-High virus to the cells led to inefficient virus internalization, low virus infectivity and reduced virus replication in a macrophage cell line. P78-High virus and P78-KO virus showed lower and higher virulence than MP-12, respectively, in young mice. ZH501-P78-High virus also exhibited lower virulence than ZH501 in mice. These data suggest that high levels of P78 expression attenuate RVFV virulence by preventing efficient virus replication in macrophages. Genetic alteration leading to increased P78 expression may serve as a novel strategy for the attenuation of RVFV virulence and generation of safe RVFV vaccines. Rift Valley fever virus (RVFV) is the causative agent of Rift Valley fever, which is primarily endemic in African countries. The virus infects a wide variety of mammalian species, including ruminants and humans, and causes devastating effects on public health and the livestock industry. Because RVFV is transmitted by several mosquito species that are ubiquitous in non-endemic areas, there is a potential risk that the virus will spread outside of the current endemic areas. However, there are no effective therapeutics nor commercially available vaccines for human use. RVFV encodes a 78kDa protein (P78) of unidentified biological function in mammalian cells. We found that a mutant RVFV expressing a higher level of P78 (P78-High virus) showed lower infectivity and less efficient replication than parental RVFV in macrophage cell lines. P78-High virus and a mutant RVFV lacking P78 expression showed lower and higher virulence than the parental virus, respectively, in mice, suggesting that high levels of P78 expression prevent efficient virus replication in macrophages, leading to attenuation of virus virulence. Genetic alteration leading to increased P78 expression may serve as a novel strategy for the generation of attenuated RVFV as vaccine candidates.
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