Virus-like particles of SARS-like coronavirus formed by membrane proteins from different origins demonstrate stimulating activity in human dendritic cells.
Virus-like particles of SARS-like coronavirus formed by membrane proteins from different origins demonstrate stimulating activity in human dendritic cells.
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DOI：
10.1371/journal.pone.0002685
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发表时间：
2008-07-16
期刊：
影响因子：
3.7
通讯作者：
Wang, Hanzhong
中科院分区：
文献类型：
--
作者：
Bai, Bingke;Hu, Qinxue;Hu, Hui;Zhou, Peng;Shi, Zhengli;Meng, Jin;Lu, Baojing;Huang, Yi;Mao, Panyong;Wang, Hanzhong
SARS冠状病毒（CoV）的发病机制仍然知之甚少。本研究制备了两种重组杆状病毒，分别表达蝙蝠分离的sars样冠状病毒（SL-CoV）的刺突(S)蛋白和SARS-CoV的包膜(E)和膜(M)蛋白。电镜显示，这两种重组杆状病毒共同感染昆虫细胞导致病毒样颗粒（BVLPs）的自组装。western blot和免疫金标法证实了vaccbs （BS）中S蛋白的掺入。这些BVLPs上调了未成熟树突状细胞（DCs）中CD40、CD80、CD86、CD83的水平，并增强了IL-6、IL-10和TNF-α的分泌。比较了未成熟dc分别接种BVLPs和由sars冠状病毒S、E和M蛋白形成的VLPs的免疫应答。BVLPs在细胞因子诱导方面表现出更强的刺激dc的能力，其IL-6和TNF-α的产量高出2至6倍。进一步的研究表明，与预先暴露于BVLPs或SARS-CoV VLPs的dc共培养后，CD4+ T细胞中的IFN-γ+和IL-4+数量增加。BVLPs和SARS冠状病毒VLPs在dc刺激活性上的差异很可能是由S蛋白引起的。与此一致的是，SL-CoV S DNA疫苗在小鼠中比SARS-CoV S DNA引起更强烈的抗体反应和更强的T细胞反应。我们的数据首次证明，由不同来源的膜蛋白组成的SL-CoV VLPs，一个来自蝙蝠的SL-CoV (BS)，另外两个来自人类SARS-CoV (E和M)，激活未成熟的dc，并增强共刺激分子的表达和细胞因子的分泌。本研究的发现可能为疫苗开发以及了解sars样冠状病毒的发病机制提供重要信息。
The pathogenesis of SARS coronavirus (CoV) remains poorly understood. In the current study, two recombinant baculovirus were generated to express the spike (S) protein of SARS-like coronavirus (SL-CoV) isolated from bats (vAcBS) and the envelope (E) and membrane (M) proteins of SARS-CoV, respectively. Co-infection of insect cells with these two recombinant baculoviruses led to self-assembly of virus-like particles (BVLPs) as demonstrated by electron microscopy. Incorporation of S protein of vAcBS (BS) into VLPs was confirmed by western blot and immunogold labeling. Such BVLPs up-regulated the level of CD40, CD80, CD86, CD83, and enhanced the secretion of IL-6, IL-10 and TNF-α in immature dendritic cells (DCs). Immune responses were compared in immature DCs inoculated with BVLPs or with VLPs formed by S, E and M proteins of human SARS-CoV. BVLPs showed a stronger ability to stimulate DCs in terms of cytokine induction as evidenced by 2 to 6 fold higher production of IL-6 and TNF-α. Further study indicated that IFN-γ+ and IL-4+ populations in CD4+ T cells increased upon co-cultivation with DCs pre-exposed with BVLPs or SARS-CoV VLPs. The observed difference in DC-stimulating activity between BVLPs and SARS CoV VLPs was very likely due to the S protein. In agreement, SL-CoV S DNA vaccine evoked a more vigorous antibody response and a stronger T cell response than SARS-CoV S DNA in mice. Our data have demonstrated for the first time that SL-CoV VLPs formed by membrane proteins of different origins, one from SL-CoV isolated from bats (BS) and the other two from human SARS-CoV (E and M), activated immature DCs and enhanced the expression of co-stimulatory molecules and the secretion of cytokines. Finding in this study may provide important information for vaccine development as well as for understanding the pathogenesis of SARS-like CoV.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
影响因子：
5.4
作者：
Buonaguro, L.;Tornesello, M. L.;Buonaguro, F. M.
通讯作者：
Buonaguro, F. M.
影响因子：
5.4
作者：
Babcock, GJ;Esshaki, DJ;Ambrosino, DM
通讯作者：
Ambrosino, DM
DOI：
10.1164/rccm.200306-766oc
发表时间：
2003-12-15
影响因子：
24.7
作者：
Ho, JC;Ooi, GC;Tsang, KW
通讯作者：
Tsang, KW
影响因子：
5.4
作者：
Corse, E;Machamer, CE
通讯作者：
Machamer, CE