Identification of Alpha Interferon-Induced Genes Associated with Antiviral Activity in Daudi Cells and Characterization of IFIT3 as a Novel Antiviral Gene
Identification of Alpha Interferon-Induced Genes Associated with Antiviral Activity in Daudi Cells and Characterization of IFIT3 as a Novel Antiviral Gene
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DOI：
10.1128/jvi.00818-10
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发表时间：
2010-10-01
影响因子：
5.4
通讯作者：
Zoon, K. C.
中科院分区：
文献类型：
--
作者：
Schmeisser, H.;Mejido, J.;Zoon, K. C.
建立了一种新的检测Daudi细胞的方法，可以同时检测干扰素（IFN）的抗病毒（AV）和抗增殖（AP）活性。利用这种新方法，确定并选择了允许IFN AV保护但不抑制生长的条件。在这些条件下处理Daudi细胞，并进行基因表达微阵列分析。分析结果确定了25个与ifn - α - AV活性相关的基因。通过逆转录- pcr证实了23个ifn诱导基因的上调。在25个基因产物中，有17个在24小时时通过Western blotting检测到。在25个基因中，有10个先前未与ifn - α的AV活性相关。上调最多的基因是IFIT3（用于ifn诱导的具有四肽重复3的蛋白）。抗体中和实验的结果表明所鉴定的基因与ifn - α AV活性有关。IFIT3小干扰RNA转染实验结果显示IFIT3表达降低，ifn - α诱导的AV活性降低，这种关联得到了加强。IFIT3过表达导致病毒滴度降低。高浓度IFN对Daudi细胞具有AP效应，经细胞处理后的AV基因转录表明鉴定的基因产物具有多效性。
A novel assay was developed for Daudi cells in which the antiviral (AV) and antiproliferative (AP) activities of interferon (IFN) can be measured simultaneously. Using this novel assay, conditions allowing IFN AV protection but no growth inhibition were identified and selected. Daudi cells were treated under these conditions, and gene expression microarray analyses were performed. The results of the analysis identified 25 genes associated with IFN-alpha AV activity. Upregulation of 23 IFN-induced genes was confirmed by using reverse transcription-PCR. Of 25 gene products, 17 were detected by Western blotting at 24 h. Of the 25 genes, 10 have not been previously linked to AV activity of IFN-alpha. The most upregulated gene was IFIT3 (for IFN-induced protein with tetratricopeptide repeats 3). The results from antibody neutralizing experiments suggested an association of the identified genes with IFN-alpha AV activity. This association was strengthened by results from IFIT3-small interfering RNA transfection experiments showing decreased expression of IFIT3 and a reduction in the AV activity induced by IFN-alpha. Overexpression of IFIT3 resulted in a decrease of virus titer. Transcription of AV genes after the treatment of cells with higher concentrations of IFN having an AP effect on Daudi cells suggested pleiotropic functions of identified gene products.