Transcriptional response of immune-related genes after endogenous expression of VP1 and exogenous exposure to VP1-based VLPs and CPV virions in lepidopteran cell lines
Transcriptional response of immune-related genes after endogenous expression of VP1 and exogenous exposure to VP1-based VLPs and CPV virions in lepidopteran cell lines
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鳞翅目细胞系中内源表达 VP1 以及外源暴露于基于 VP1 的 VLP 和 CPV 病毒颗粒后免疫相关基因的转录反应
DOI：
10.1007/s00438-019-01551-1
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发表时间：
2019-03
影响因子：
3.1
通讯作者：
Sun Jingchen
中科院分区：
文献类型：
--
作者：
Zhao Yongchao;Kolliopoulou Anna;Ren Feifei;Lu Qiuyuan;Labropoulou Vassiliki;Swevers Luc;Sun Jingchen
在昆虫中，RNAi被认为是主要的抗病毒免疫防御途径。在病毒感染期间产生的DsRNA被Dicer酶加工成小RNA，其作为特异性决定子来沉默病毒基因。相比之下，在哺乳动物中，模式识别受体（PRR）对与病毒感染相关的分子（例如dsRNA）的识别启动信号级联，其最终产生和释放具有抗病毒功能的信号蛋白（例如干扰素）。然而，在昆虫中，病毒体的组分可以被识别为病原体激活的分子模式（PAMPs）来激活先天免疫应答的假说尚未得到系统的研究。在这项研究中，潜在的VP 1，构成的主要衣壳蛋白质的胞质型多角体病毒（CPV;呼肠孤病毒科），激活一个集合的免疫相关基因进行了检查，在家蚕衍生的Bm 5细胞。测试了两种不同的VP 1给药方法，通过转化细胞系中的内源性表达，或通过将纯化的基于VP 1的病毒样颗粒添加到细胞外培养基中。此外，还进行了暴露于从纯化的多角体分离的CPV病毒体。总的来说，我们的研究结果没有显示免疫相关基因对VP 1或CPV病毒粒子的强转录应答，但注意到两个例外。首先，抗菌肽（AMP）基因Attacin的表达强烈诱导后24小时暴露于基于VP 1的VLP。第二，在表达VP 1的转化细胞Sf 21中，dcr-2（RNAi途径中的必需基因）的表达水平显著增加，而在转化细胞Bm 5中没有，表明存在物种特异性效应。然而，当在RNAi报告基因测定中测试时，dcr-2表达的增加并没有导致沉默效率的增加。我们的研究表明，CPV的衣壳蛋白VP 1具有作为PAMP的潜力，并在昆虫细胞中诱导与RNAi和蛋白质效应物如AMP相关的转录反应。PRR的身份和信号级联，可能是由VP 1触发的，仍有待于在未来的实验中阐明。虽然这项研究是在小规模上进行的，但它可以鼓励使用高通量方法（微阵列，深度测序）进行更全面的研究，以更系统地搜索病毒衣壳蛋白是否可以在昆虫中充当PAMP，以及它们的产生是否会导致诱导具有潜在抗病毒功能的免疫相关基因。
In insects, RNAi is considered the major antiviral immune defense pathway. DsRNAs produced during viral infection are processed by Dicer enzymes into small RNAs that function as specificity determinants to silence viral genes. By contrast, in mammals, recognition of molecules associated with viral infection, such as dsRNA, by pattern recognition receptors (PRRs) initiates a signaling cascade that culminates in the production and release of signaling proteins with antiviral function such as interferons. However, in insects, the hypothesis that components of virions can be recognized as pathogen-activated molecular patterns (PAMPs) to activate the innate immune response has not been investigated systematically. In this study, the potential of VP1, that constitutes the major capsid protein of cytoplasmic polyhedrosis virus (CPV; Reoviridae), to activate a collection of immune-related genes was examined in silkworm-derived Bm5 cells. Two different methods of VP1 administration were tested, either through endogenous expression in transformed cell lines, or through addition of purified VP1-based viral-like particles to the extracellular medium. In addition, exposure to CPV virions isolated from purified polyhedra was also performed. In general, our results do not show a robust transcriptional response of immune-related genes to VP1 or CPV virions, but two exceptions were noted. First, the expression of the antimicrobial peptide (AMP) geneAttacinwas strongly induced after 24 h of exposure to VP1-based VLPs. Second, the expression levels ofdcr-2, an essential gene in the RNAi pathway, were greatly increased in VP1-expressing transformed Sf21 cells but not transformed Bm5 cells, indicating the existence of species-specific effects. However, the increased expression ofdcr-2did not result in increased silencing efficiency when tested in an RNAi reporter assay. Our study indicates that the capsid protein VP1 of CPV has the potential to act as a PAMP and to induce a transcriptional response in insect cells that relate both to RNAi and protein effectors such as AMPs. The identity of the PRRs and the signaling cascade that are potentially triggered by VP1 remain to be elucidated in future experiments. While this study was performed on a small scale, it can encourage more comprehensive studies with high-throughput approaches (microarray, deep sequencing) to search more systematically whether viral capsid proteins can act as PAMPs in insects and whether their production results in the induction of immune-related genes with potential antiviral function.
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影响因子：
3.8
作者：
Hagiwara, K;Rao, SJ;Carner, GR
通讯作者：
Carner, GR
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影响因子：
3.1
作者：
Qi Qi;Yao Lunguang;Liang Zhisheng;Yan Donghua;Li Zhuo;Huang Yadong;Sun Jingchen
通讯作者：
Sun Jingchen
影响因子：
5.6
作者：
Kingsolver MB;Huang Z;Hardy RW
通讯作者：
Hardy RW
影响因子：
2.2
作者：
L. Swevers;Anna Kolliopoulou;Zheng Li;Maria G Daskalaki;Frédéric Verret;K. Kalantidis;G. Smagghe;Jingchen Sun
通讯作者：
L. Swevers;Anna Kolliopoulou;Zheng Li;Maria G Daskalaki;Frédéric Verret;K. Kalantidis;G. Smagghe;Jingchen Sun
影响因子：
4.6
作者：
Merkling SH;Overheul GJ;van Mierlo JT;Arends D;Gilissen C;van Rij RP
通讯作者：
van Rij RP