Xin Liu, Xueying Song, Xiaoping Zhang, Ying-lun Han, Ting Zhu, R. Xiao, Qing-wei Li
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引用次数: 2
摘要
近年来在七鳃鳗中发现了基于可变淋巴细胞受体(VLRs)的抗原识别机制。为了阐明七鳃鳗粘膜免疫系统中淋巴细胞样细胞免疫应答的遗传基础,探索无颌和有颌脊椎动物之间适应性免疫应答的进化关系,我们构建了刺激前后七鳃鳗(Lampetra japonica)鳃的cDNA文库,并进行了高通量转录组测序和分析。通过对88 525个组装的unigenes进行功能标注,分别在Gene Ontology (GO)和Kyto Encyclopedia of Genes and Genomes (KEGG)数据库中标注了21 704个和9769个unigenes。在参与免疫系统多通路的999个unique igenes中,184个unique igenes与51个高等脊椎动物的TCR (T细胞受体)和BCR (B细胞受体)信号分子高度同源,说明参与高等脊椎动物适应性免疫信号通路的分子在七鳃鳗中也存在。此外,5个VLRA、7个VLRB和4个VLRC分子的鉴定表明,在七鳃鳗鳃黏膜免疫组织中至少分布着3种类型的淋巴细胞亚群。实时荧光定量PCR结果显示,免疫刺激后Lck、Fyn和Zap70的表达水平上调,而Syk、Btk和Blnk的表达水平变化不明显,表明抗原刺激后七鳃鱼鳃组织中tcr样信号转导通路被激活。我们的研究初步证明了无颌和有颌脊椎动物两种平行的适应性免疫系统具有共同的遗传基础,也为探索VLRA +、VLRB +、VLRC +淋巴细胞样细胞对抗原反应的信号传导过程提供了有价值的线索。
[Genetic basis of immune response of lymphocyte-like cells in the mucosal immune system of Lampetra japonica].
In recent years, the antigen recognition mechanism based on variable lymphocyte receptors (VLRs) was found in agnathan lamprey. To illuminate the genetic basis of immune response of lymphocyte-like cells in the mucosal immune system of lamprey and explore the evolutionary relationship of adaptive immune responses between the jawless and jawed vertebrates, we constructed cDNA libraries of lamprey (Lampetra japonica) gills before and after stimulation, and then performed high-throughput transcriptome sequencing and analysis. Through functional annotation of 88 525 assembled unigenes, 21 704 and 9769 unigenes were annotated in Gene Ontology (GO) and Kyto Encyclopedia of Genes and Genomes (KEGG) databases, respectively. Among 999 unigenes involved in multiple pathways of immune system, 184 unigenes were highly homologous to 51 TCR (T cell receptor) and BCR (B cell receptor) signalling molecules in higher vertebrates, indicating that molecules involved in adaptive immune signalling pathways in higher vertebrates also exist in lampreys. In addition, identification of five VLRA, seven VLRB and four VLRC molecules suggest that at least three types of lymphocyte subsets are distributed in lamprey gill mucosal immune tissues. The results of real-time fluorescence quantitative PCR showed that the expression levels of Lck, Fyn and Zap70 were up-regulated after immune stimulation while those of Syk, Btk and Blnk were not changed significantly, indicating the activation of TCR-like signal transduction pathway after antigen stimulation in lamprey gill tissues. Our studies preliminaryly proved that two parallel adaptive immune systems in jawless and jawed vertebrates have common genetic basis, and also provided valuable clues to the exploration of signalling processes of VLRA⁺, VLRB⁺, and VLRC⁺ lymphocyte-like cells in response to antigens.