SARS coronavirus spike protein-induced innate immune response occurs via activation of the NF-kappaB pathway in human monocyte macrophages in vitro.
SARS coronavirus spike protein-induced innate immune response occurs via activation of the NF-kappaB pathway in human monocyte macrophages in vitro.
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DOI：
10.1016/j.virusres.2009.01.005
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发表时间：
2009-06
期刊：
影响因子：
5
通讯作者：
Collins, Arlene R.
中科院分区：
文献类型：
--
作者：
Dosch, Susan F.;Mahajan, Supriya D.;Collins, Arlene R.
研究了纯化的重组刺突 (S) 蛋白对刺激人外周血单核巨噬细胞 (PBMC) 的影响。我们使用基因阵列检查了 S 蛋白处理的 PBMC 中发现的炎症基因 mRNA 丰度。我们鉴定了具有与抗病毒（CXCL10）和炎症（IL-6 和 IL-8）反应相关的功能特性的基因的差异 mRNA 丰度。我们通过实时定量 (q) RT-PCR 或 ELISA 证实了细胞因子 mRNA 的增加。我们进一步分析了突出的 IL-8 反应的敏感性和特异性。通过实时 qRT-PCR，S 蛋白以剂量依赖性方式刺激 IL-8 mRNA 积累，而 E 蛋白处理则不然。此外，通过 ELISA 对 S 蛋白特异性产生和 IL-8 分泌的滴定表明，与模拟处理对照相比，5.6 nM S 剂量可显着增加 IL-8 (p = 0.003)。 5.6 nM S 浓度刺激的 IL-8 增加与 S 蛋白与 ACE2 或中和单克隆抗体结合的浓度相当，表明存在生理相关性。 TPCK（N-Tosyl-L-Phenylalanine Chloromethyl Ketone）是一种 NF-κB 抑制剂，可以抑制 PBMC 和 THP-1 细胞以及 HCoV-229E 病毒感染的 PBMC 中响应 S 蛋白的 IL-8 产生和分泌。使用高灵敏度的活性核 NF-κB p65 转录因子测定法显示，PBMC 或 THP-1 细胞暴露于 S 蛋白后，NF-κB 的激活和易位迅速发生。结果进一步表明，释放或分泌的S蛋白可以通过Toll样受体（TLR）2配体的识别来激活血液单核细胞。
A purified recombinant spike (S) protein was studied for its effect on stimulating human peripheral blood monocyte macrophages (PBMC). We examined inflammatory gene mRNA abundances found in S protein-treated PBMC using gene arrays. We identified differential mRNA abundances of genes with functional properties associated with antiviral (CXCL10) and inflammatory (IL-6 and IL-8) responses. We confirmed cytokine mRNA increases by real-time quantitative(q) RT-PCR or ELISA. We further analyzed the sensitivity and specificity of the prominent IL-8 response. By real-time qRT-PCR, S protein was shown to stimulate IL-8 mRNA accumulation in a dose dependent manner while treatment with E protein did not. Also, titration of S protein-specific production and secretion of IL-8 by ELISA showed that the dose of 5.6 nM of S produced a significant increase in IL-8 (p = 0.003) compared to mock-treated controls. The increase in IL-8 stimulated by a concentration of 5.6 nM of S was comparable to concentrations seen for S protein binding to ACE2 or to neutralizing monoclonal antibody suggesting a physiological relevance. An NF-κB inhibitor, TPCK (N-Tosyl-L-Phenylalanine Chloromethyl Ketone) could suppress IL-8 production and secretion in response to S protein in PBMC and THP-1 cells and in HCoV-229E virus-infected PBMC. Activation and translocation of NF-κB was shown to occur rapidly following exposure of PBMC or THP-1 cells to S protein using a highly sensitive assay for active nuclear NF-κB p65 transcription factor. The results further suggested that released or secreted S protein could activate blood monocytes through recognition by toll-like receptor (TLR)2 ligand.
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DOI：
10.1074/jbc.m508381200
发表时间：
2006-02-10
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Huang IC;Bosch BJ;Li F;Li W;Lee KH;Ghiran S;Vasilieva N;Dermody TS;Harrison SC;Dormitzer PR;Farzan M;Rottier PJ;Choe H
通讯作者：
Choe H
影响因子：
7.4
作者：
Davis, JN;Kucuk, O;Sarkar, FH
通讯作者：
Sarkar, FH
影响因子：
7.3
作者：
He, L.;Ding, Y.;Jiang, S.
通讯作者：
Jiang, S.
影响因子：
3.1
作者：
Elass, E;Aubry, L;Kremer, L
通讯作者：
Kremer, L
影响因子：
5.4
作者：
Bosch, Berend Jan;Bartelink, Willem;Rottier, Peter J. M.
通讯作者：
Rottier, Peter J. M.