Callol Agnès, Reyes-López Felipe E, Roig Francisco J, Goetz Giles, Goetz Frederick W, Amaro Carmen, MacKenzie Simon A
Departament de Microbiologia i Ecologia, Universitat de Valencia, Burjassot, Spain; Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, Spain; Department de Biologia cel·lular, Fisiologia Animal i Immunologia, Universitat Autònoma de Barcelona, Bellaterra, Spain.
PLoS One. 2015 Jul 24;10(7):e0133328. doi: 10.1371/journal.pone.0133328. eCollection 2015.
Infectious diseases are one of the principal bottlenecks for the European eel recovery. The aim of this study was to develop a new molecular tool to be used in host-pathogen interaction experiments in the eel. To this end, we first stimulated adult eels with different pathogen-associated molecular patterns (PAMPs), extracted RNA from the immune-related tissues and sequenced the transcriptome. We obtained more than 2 x 10(6) reads that were assembled and annotated into 45,067 new descriptions with a notable representation of novel transcripts related with pathogen recognition, signal transduction and the immune response. Then, we designed a DNA-microarray that was used to analyze the early immune response against Vibrio vulnificus, a septicemic pathogen that uses the gills as the portal of entry into the blood, as well as the role of the main toxin of this species (RtxA13) on this early interaction. The gill transcriptomic profiles obtained after bath infecting eels with the wild type strain or with a mutant deficient in rtxA13 were analyzed and compared. Results demonstrate that eels react rapidly and locally against the pathogen and that this immune-response is rtxA13-dependent as transcripts related with cell destruction were highly up-regulated only in the gills from eels infected with the wild-type strain. Furthermore, significant differences in the immune response against the wild type and the mutant strain also suggest that host survival after V. vulnificus infection could depend on an efficient local phagocytic activity. Finally, we also found evidence of the presence of an interbranchial lymphoid tissue in European eel gills although further experiments will be necessary to identify such tissue.
传染病是欧洲鳗鲡恢复的主要瓶颈之一。本研究的目的是开发一种新的分子工具,用于鳗鲡宿主-病原体相互作用实验。为此,我们首先用不同的病原体相关分子模式(PAMP)刺激成年鳗鲡,从免疫相关组织中提取RNA并对转录组进行测序。我们获得了超过2×10⁶条读数,这些读数被组装并注释为45,067个新的描述,其中有大量与病原体识别、信号转导和免疫反应相关的新转录本。然后,我们设计了一种DNA微阵列,用于分析针对创伤弧菌的早期免疫反应,创伤弧菌是一种通过鳃作为进入血液门户的败血性病原体,以及该物种主要毒素(RtxA13)在这种早期相互作用中的作用。分析并比较了用野生型菌株或rtxA13缺陷型突变体对鳗鲡进行浸浴感染后获得的鳃转录组图谱。结果表明,鳗鲡对病原体反应迅速且具有局部性,并且这种免疫反应依赖于rtxA13,因为与细胞破坏相关的转录本仅在感染野生型菌株的鳗鲡的鳃中高度上调。此外,针对野生型和突变型菌株的免疫反应的显著差异也表明,创伤弧菌感染后宿主的存活可能取决于有效的局部吞噬活性。最后,我们还发现了欧洲鳗鲡鳃中存在鳃间淋巴组织的证据,尽管还需要进一步的实验来鉴定这种组织。