SARS-CoV-2 vaccine ChAdOx1 nCoV-19 infection of human cell lines reveals low levels of viral backbone gene transcription alongside very high levels of SARS-CoV-2 S glycoprotein gene transcription.
SARS-CoV-2 vaccine ChAdOx1 nCoV-19 infection of human cell lines reveals low levels of viral backbone gene transcription alongside very high levels of SARS-CoV-2 S glycoprotein gene transcription.
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DOI：
10.1186/s13073-021-00859-1
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发表时间：
2021-03-15
期刊：
影响因子：
12.3
通讯作者：
Matthews DA
中科院分区：
文献类型：
--
作者：
Almuqrin A;Davidson AD;Williamson MK;Lewis PA;Heesom KJ;Morris S;Gilbert SC;Matthews DA
Chadox1 NCOV-19是一种针对SARS-COV-2的重组腺病毒疫苗，该疫苗已通过III期临床试验，目前正在全球使用，尽管在正常细胞中的复制缺陷，但腺病毒基因的28 kbp被递送到细胞核中与SARS-COV-2 S糖蛋白基因一起。
我们使用直接的RNA测序来分析人类MRC-5的Chadox1 NCOV-19基因组和A549细胞系中的转录物表达，这些细胞系与复制允许的细胞系一起使用，这些载体复制与载体复制，另外，我们还将定量蛋白质组学用于定量蛋白质组。随着时间的流逝，A549和MRC5细胞感染了Chadox1 NCOV-19疫苗的蛋白质组和磷酸蛋白酶。
预期的SARS-COV-2 S编码在所有细胞系中都占主导地位。除S糖蛋白外，我们还检测到A549细胞中多种腺病毒蛋白的腺病毒基因表达的更广泛的曲目在MRC5细胞中只有一个。
总体而言，Chadox1 NCOV-19疫苗在细胞培养中的转录组和蛋白质组学库是预期的。该技术为未来的病毒疫苗媒介设计提供了潜力。
在线版本中包含10.1186/s13073-021-00859-1的补充材料。
ChAdOx1 nCoV-19 is a recombinant adenovirus vaccine against SARS-CoV-2 that has passed phase III clinical trials and is now in use across the globe. Although replication-defective in normal cells, 28 kbp of adenovirus genes is delivered to the cell nucleus alongside the SARS-CoV-2 S glycoprotein gene. We used direct RNA sequencing to analyse transcript expression from the ChAdOx1 nCoV-19 genome in human MRC-5 and A549 cell lines that are non-permissive for vector replication alongside the replication permissive cell line, HEK293. In addition, we used quantitative proteomics to study over time the proteome and phosphoproteome of A549 and MRC5 cells infected with the ChAdOx1 nCoV-19 vaccine. The expected SARS-CoV-2 S coding transcript dominated in all cell lines. We also detected rare S transcripts with aberrant splice patterns or polyadenylation site usage. Adenovirus vector transcripts were almost absent in MRC-5 cells, but in A549 cells, there was a broader repertoire of adenoviral gene expression at very low levels. Proteomically, in addition to S glycoprotein, we detected multiple adenovirus proteins in A549 cells compared to just one in MRC5 cells. Overall, the ChAdOx1 nCoV-19 vaccine’s transcriptomic and proteomic repertoire in cell culture is as expected. The combined transcriptomic and proteomics approaches provide a detailed insight into the behaviour of this important class of vaccine using state-of-the-art techniques and illustrate the potential of this technique to inform future viral vaccine vector design. The online version contains supplementary material available at 10.1186/s13073-021-00859-1.
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影响因子：
5.5
作者：
Bangari, DS;Mittal, SK
通讯作者：
Mittal, SK
影响因子：
5.9
作者：
Donovan-Banfield, I'ah;Turnell, Andrew S.;Matthews, David A.
通讯作者：
Matthews, David A.
影响因子：
5.4
作者：
Lusky, M;Christ, M;Mehtali, M
通讯作者：
Mehtali, M
DOI：
10.1038/mt.2013.284
发表时间：
2014-03
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.8
作者：
GRAHAM, FL;SMILEY, J;NAIRN, R
通讯作者：
NAIRN, R