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Transcriptome analysis of Ehrlichia ruminantium in the ruminant host at the tick bite site and in the tick vector salivary glands

机译:蜱咬伤位点的反刍动物宿主中Ehrlichia强膜的转录组分析及蜱瘤唾液腺

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摘要

Heartwater is a non-contagious tick-borne disease of domestic and wild ruminants. Data regarding the complex processes involved during pathogen-vector?host interaction during Ehrlichia ruminantium infection is lacking and could be improved with knowledge associated with gene expression changes in both the pathogen and the host. Thus, in the current study, we aimed to identify E. ruminantium genes that are up-regulated when the pathogen enters the host and before the disease is established. Identification of such genes/proteins may aid in future vaccine development strategies against heartwater. RNA-sequencing was used to identify E. ruminantium genes that were exclusively expressed at the tick bite site in sheep skin biopsies (SB) and in adult tick salivary glands (SG). RNA was extracted from pooled samples of the SB or SG collected at different time points during tick attachment and prior to disease manifestation. Ribosomal RNA (rRNA) was removed and the samples were sequenced. Several E. ruminantium genes were highly expressed in all the samples while others were exclusively expressed in each. It was concluded that E. ruminantium genes that were exclusively expressed in the SB or both SB and SG when compared to the transcriptome datasets from bovine elementary bodies (BovEBs) from cell culture may be considered as early antigenic targets of host immunity. In silico immunogenic epitope prediction analysis and preliminary characterization of selected genes in vitro using ELIspot assay showed that they could possibly be ideal targets for future vaccine development against heartwater, however, further epitope characterization is still required.
机译:心脏水是家养和野生反刍动物的一种非传染性蜱传播疾病。关于病原体传播过程中涉及的复杂过程的数据?反刍埃立克体感染期间缺乏宿主相互作用,可以通过了解病原体和宿主的基因表达变化来改善这种相互作用。因此,在目前的研究中,我们的目的是鉴定在病原体进入宿主和疾病形成之前上调的反刍动物基因。识别这些基因/蛋白质可能有助于未来针对心脏水的疫苗开发策略。RNA测序用于鉴定反刍动物E.ruminatium基因,这些基因在绵羊皮肤活检(SB)和成年蜱唾液腺(SG)中仅在蜱叮咬部位表达。RNA是从蜱虫附着期间和疾病表现之前不同时间点收集的SB或SG样本中提取的。去除核糖体RNA(rRNA)并对样本进行测序。几个反刍动物基因在所有样本中高度表达,而其他基因在每个样本中都只表达。结论是,与细胞培养的牛基本体(BovEBs)转录组数据集相比,仅在SB或SB和SG中表达的反刍动物E.基因可被视为宿主免疫的早期抗原靶标。电子免疫原性表位预测分析和体外ELIspot分析对所选基因的初步鉴定表明,它们可能是未来心脏水疫苗开发的理想靶点,但仍需进一步的表位鉴定。

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