绵羊T淋巴细胞对人工初次捻转血矛线虫感染反应中差异表达基因的分析
Profiling of differentially expressed genes in sheep T lymphocytes response to an artificial primary Haemonchus contortus infection.
作者信息
Yang Yi, Zhou Qian-Jin, Chen Xue-Qiu, Yan Bao-Long, Guo Xiao-Lu, Zhang Hong-Li, Du Ai-Fang
机构信息
Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China.
出版信息
Parasit Vectors. 2015 Apr 18;8:235. doi: 10.1186/s13071-015-0844-z.
BACKGROUND
Haemonchus contortus is a common bloodsucking nematode causing widespread economic loss in agriculture. Upon H. contortus infection, a series of host responses is elicited, especially those related to T lymphocyte immunity. Existing studies mainly focus on the general immune responses of sheep T lymphocyte to H. contortus, lacking investigations at the molecular level. The objective of this study was to obtain a systematic transcriptional profiling of the T lymphocytes in H. contortus primary-infected sheep.
METHODS
Nematode-free sheep were orally infected once with H. contortus L3s. T lymphocyte samples were collected from the peripheral blood of 0, 3, 30 and 60 days post infection (dpi) infected sheep. Microarrays were used to compare gene transcription levels between samples. Quantitative RT-PCR was employed to validate the microarray data. Gene Ontology and KEGG pathway analysis were utilized for the annotation of differentially expressed genes.
RESULTS
Our microarray data was consistent with qPCR results. From microarrays, 853, 242 and 42 differentially expressed genes were obtained in the 3d vs. 0d, 30d vs. 0d and 60d vs. 0d comparison groups, respectively. Gene Ontology and KEGG pathway analysis indicated that these genes were involved in metabolism, signaling, cell growth and immune system processes. Functional analysis of significant differentially expressed genes, such as SLC9A3R2, ABCB9, COMMD4, SUGT1, FCER1G, GSK3A, PAK4 and FCER2, revealed a crucial association with cellular homeostasis maintenance and immune response. Our data suggested that maintaining both effective immunological response and natural cellular activity are important for T lymphocytes in fighting against H. contortus infection.
CONCLUSIONS
Our results provide a substantial list of candidate genes in sheep T lymphocytes response to H. contortus infection, and contribute novel insights into a general immune response upon infection.
背景
捻转血矛线虫是一种常见的吸血线虫，在农业中造成广泛的经济损失。感染捻转血矛线虫后，会引发一系列宿主反应，尤其是那些与T淋巴细胞免疫相关的反应。现有研究主要集中于绵羊T淋巴细胞对捻转血矛线虫的一般免疫反应，缺乏分子水平的研究。本研究的目的是获得捻转血矛线虫初次感染绵羊中T淋巴细胞的系统转录谱。
方法
无线虫绵羊经口一次性感染捻转血矛线虫L3期幼虫。在感染后0、3、30和60天（dpi）从感染绵羊的外周血中收集T淋巴细胞样本。使用微阵列比较样本之间的基因转录水平。采用定量RT-PCR验证微阵列数据。利用基因本体论和KEGG通路分析对差异表达基因进行注释。
结果
我们的微阵列数据与qPCR结果一致。通过微阵列，在3d vs. 0d、30d vs. 0d和60d vs. 0d比较组中分别获得了853、242和42个差异表达基因。基因本体论和KEGG通路分析表明，这些基因参与代谢、信号传导、细胞生长和免疫系统过程。对显著差异表达基因（如SLC9A3R2、ABCB9、COMMD4、SUGT1、FCER1G、GSK3A、PAK4和FCER2）的功能分析揭示了其与细胞稳态维持和免疫反应的关键关联。我们的数据表明，维持有效的免疫反应和自然细胞活性对于T淋巴细胞抵抗捻转血矛线虫感染都很重要。
结论
我们的结果提供了绵羊T淋巴细胞对捻转血矛线虫感染反应中的大量候选基因列表，并为感染后的一般免疫反应提供了新的见解。
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