A simian virus 5 (SV5) P/V mutant is less cytopathic than wild-type SV5 in human dendritic cells and is a more effective activator of dendritic cell maturation and function
A simian virus 5 (SV5) P/V mutant is less cytopathic than wild-type SV5 in human dendritic cells and is a more effective activator of dendritic cell maturation and function
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DOI：
10.1128/jvi.80.7.3416-3427.2006
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发表时间：
2006-04-01
影响因子：
5.4
通讯作者：
Parks, GD
中科院分区：
文献类型：
--
作者：
Arimilli, S;Alexander-Miller, MA;Parks, GD
副流感病毒猴病毒5（SV 5）感染的人上皮细胞显示宿主细胞干扰素（IFN）、细胞因子和细胞死亡途径的最小激活。相反，重组SV 5 P/V基因突变体（rSV 5 P/V-CPI-）过表达病毒基因产物，是这些细胞中IFN、促炎细胞因子和凋亡的有效诱导剂。在这项研究中，我们比较了野生型（WT）SV 5和rSV 5-P/V-CPI-感染原代人树突状细胞（DC）的结果，树突状细胞是启动适应性免疫反应的重要抗原呈递细胞。我们已经测试了这样的假设，即激活宿主抗病毒反应的P/V突变体将是比抑制宿主细胞反应的WT rSV 5更有效的DC成熟和功能诱导剂。WT rSV 5感染外周血单核细胞来源的未成熟DC导致高水平的病毒蛋白和子代病毒，但细胞表面共刺激分子或IFN和促炎细胞因子的分泌很少增加。相反，用rSV 5-P/V-CPI-突变体感染的未成熟DC仅产生低水平的病毒蛋白和子代病毒，但这些感染的细胞被诱导分泌IFN-α和其他细胞因子，并显示出升高水平的成熟标志物。出乎意料的是，感染WT rSV 5的DC表现出广泛的细胞病变效应和活性caspase-3水平的增加，而感染P/V突变体的DC在很大程度上是非细胞病变的。在混合培养试验中，WT rSV 5感染的DC刺激自体CD 4（+）T细胞增殖的能力受损，而P/V突变体感染的DC在激活T细胞增殖方面非常有效。向用WT rSV 5感染的DC中加入泛胱天蛋白酶抑制剂减少了细胞病变效应，并导致成熟标志物的更高表面表达水平。我们的发现，SV 5的P/V突变体有一个减少的细胞病变的影响，在人类DC相比，WT SV 5和增强的能力，诱导DC功能的合理设计的基础上，在病毒P/V基因的工程突变的新型重组副粘病毒载体的影响。
Human epithelial cells infected with the parainfluenza virus simian virus 5 (SV5) show minimal activation of host cell interferon (IFN), cytokine, and cell death pathways. In contrast, a recombinant SV5 P/V gene mutant (rSV5P/V-CPI-) overexpresses viral gene products and is a potent inducer of IFN, proinflammatory cytokines, and apoptosis in these cells. In this study, we have compared the outcomes of wild-type (WT) SV5 and rSV5-P/V-CPI- infections of primary human dendritic cells (DC), important antigen-presenting cells for initiating adaptive immune responses. We have tested the hypothesis that a P/V mutant which activates host antiviral responses will be a more potent inducer of DC maturation and function than WT rSV5, which suppresses host cell responses. Infection of peripheral blood mononuclear cell-derived immature DC with WT rSV5 resulted in high levels of viral protein and progeny virus but very little increase in cell surface costimulatory molecules or secretion of IFN and proinflammatory cytokines. In contrast, immature DC infected with the rSV5-P/V-CPI- mutant produced only low levels of viral protein and progeny virus, but these infected cells were induced to secrete IFN-alpha and other cytokines and showed elevated levels of maturation markers. Unexpectedly, DC infected with WT rSV5 showed extensive cytopathic effects and increased levels of active caspase-3, while infection of DC with the P/V mutant was largely noncytopathic. In mixed-culture assays, WT rSV5-infected DC were impaired in the ability to stimulate proliferation of autologous CD4(+) T cells, whereas DC infected with the P/V mutant were very effective at activating T-cell proliferation. The addition of a pancaspase inhibitor to DC infected with WT rSV5 reduced cytopathic effects and resulted in higher surface expression levels of maturation markers. Our finding that the SV5 P/V mutant has both a reduced cytopathic effect in human DC compared to WT SV5 and an enhanced ability to induce DC function has implications for the rational design of novel recombinant paramyxovirus vectors based on engineered mutations in the viral P/V gene.