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RNA-seq 肝脏转录组分析显示,在鱼类疫苗免疫的早期阶段,MHC-I 途径被激活,而 MHC-II 途径受到抑制。

RNA-seq liver transcriptome analysis reveals an activated MHC-I pathway and an inhibited MHC-II pathway at the early stage of vaccine immunization in zebrafish.

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.

出版信息

BMC Genomics. 2012 Jul 17;13:319. doi: 10.1186/1471-2164-13-319.

Abstract

BACKGROUND

Zebrafish (Danio rerio) is a prominent vertebrate model of human development and pathogenic disease and has recently been utilized to study teleost immune responses to infectious agents threatening the aquaculture industry. In this work, to clarify the host immune mechanisms underlying the protective effects of a putative vaccine and improve its immunogenicity in the future efforts, high-throughput RNA sequencing technology was used to investigate the immunization-related gene expression patterns of zebrafish immunized with Edwardsiella tarda live attenuated vaccine.

RESULTS

Average reads of 18.13 million and 14.27 million were obtained from livers of zebrafish immunized with phosphate buffered saline (mock) and E. tarda vaccine (WED), respectively. The reads were annotated with the Ensembl zebrafish database before differential expressed genes sequencing (DESeq) comparative analysis, which identified 4565 significantly differentially expressed genes (2186 up-regulated and 2379 down-regulated in WED; p<0.05). Among those, functional classifications were found in the Gene Ontology database for 3891 and in the Kyoto Encyclopedia of Genes and Genomes database for 3467. Several pathways involved in acute phase response, complement activation, immune/defense response, and antigen processing and presentation were remarkably affected at the early stage of WED immunization. Further qPCR analysis confirmed that the genes encoding the factors involved in major histocompatibility complex (MHC)-I processing pathway were up-regulated, while those involved in MHC-II pathway were down-regulated.

CONCLUSION

These data provided insights into the molecular mechanisms underlying zebrafish immune response to WED immunization and might aid future studies to develop a highly immunogenic vaccine against gram-negative bacteria in teleosts.

摘要

背景

斑马鱼(Danio rerio)是一种重要的脊椎动物模型,可用于研究人类发育和致病疾病,最近也被用于研究鱼类对威胁水产养殖业的传染性病原体的免疫反应。在这项工作中,为了阐明潜在疫苗保护作用的宿主免疫机制,并在未来的研究中提高其免疫原性,我们利用高通量 RNA 测序技术研究了斑马鱼用爱德华氏菌活疫苗免疫后的免疫相关基因表达模式。

结果

用磷酸盐缓冲盐水(mock)和 E. tarda 疫苗(WED)免疫的斑马鱼肝脏分别获得了 1813 万和 1427 万条平均读数。在进行差异表达基因测序(DESeq)比较分析之前,将这些读数用 Ensembl 斑马鱼数据库进行注释,共鉴定出 4565 个显著差异表达基因(WED 中 2186 个上调和 2379 个下调;p<0.05)。其中,有 3891 个在基因本体数据库中有功能分类,3467 个在京都基因与基因组百科全书数据库中有功能分类。一些参与急性期反应、补体激活、免疫/防御反应和抗原加工与呈递的途径在 WED 免疫的早期阶段受到显著影响。进一步的 qPCR 分析证实,参与 MHC-I 加工途径的因子的基因上调,而参与 MHC-II 途径的基因下调。

结论

这些数据为斑马鱼对 WED 免疫的免疫反应的分子机制提供了深入的了解,并可能有助于未来的研究,开发出针对鱼类革兰氏阴性菌的高免疫原性疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/af80/3583171/b6f2b507824b/1471-2164-13-319-1.jpg

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