| 摘要: |
| 为研究非复制型尿嘧啶营养缺陷型弓形虫（Nonreplicating Toxoplasma uracil auxotrophs，NRTUAs）在转录组水平对雏鸡胸腺的免疫调节作用，本研究以接种NRTUAs虫株3 d的雏鸡为试验对象，利用RNA-seq技术对其胸腺组织进行转录组测序以挖掘其中的差异表达基因（DEGs），进而对得到的DEGs进行GO功能注释、KEGG信号通路富集分析和蛋白互作网络分析。结果表明：1）筛选出964个DEGs，其中517个基因表达上调，447个表达下调。2）通过GO功能注释发现差异基因在生物过程、细胞组分和分子功能三大类别显著富集在免疫应答、防御反应、对外界刺激反应、对刺激反应、细胞外区域、细胞表面等条目。3）KEGG信号通路富集分析发现DEGs显著集中在细胞因子及其受体相互作用、IgA产生的肠道免疫网络、细胞粘附分子、Toll样受体信号通路等通路上。4）通过蛋白互作网络分析发现9个重要的关键分子，其中IL-6和IL-18具有调节细胞分化促进干扰素产生的功能，IRF1和IRF7能够调节巨噬细胞等细胞的活化以及抗原递呈能力，SOCS1能够防止STAT1信号通路异常活跃而导致的过度炎症反应，CCL4、CCR5和CCL20作为趋化因子能够诱导多种免疫细胞浸润炎症区域，CD40能够增强抗原递呈进而调节机体的免疫功能。综上，NRTUAs虫株刺激雏鸡胸腺后，胸腺细胞的增殖活动减弱，T细胞活化以及细胞因子分泌等活动增强，并且IL-6、IL-18、IRF1、IRF7、SOCS1、CCL4、CCR5、CCL20和CD40是NRTUAs虫株调节雏鸡胸腺细胞的重要分子，本研究为深入探讨NRTUAs虫株调节家禽胸腺的先天性免疫反应的分子机制提供了理论依据。 |
| 关键词: 非复制型尿嘧啶营养缺陷型弓形虫 转录组学 雏鸡 胸腺 |
| DOI：10.11841/j.issn.1007-4333.2024.09.11 |
| 投稿时间：2024-01-22 |
| 基金项目:国家自然科学基金项目（31902277）；中央引导地方科技发展资金科技成果转移转化项目（23ZYCGSN00080） |
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| Transcriptome analysis of chicken thymocytes stimulated by nonreplicating Toxoplasma uracil auxotrophs |
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LIAO Yanpeng, ZENG Yuhang, JING Daiyun, REN Chao
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| (Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry/College of Animal Science and Veterinary Medicine， Tianjin Agricultural University，Tianjin 300392， China) |
| Abstract: |
| In order to investigate the immunomodulatory effects of nonreplicating Toxoplasma uracil auxotrophs （NRTUAs） on the thymus of chickens at the transcriptional level， this study employed RNA-seq technology to conduct transcriptome sequencing on thymus of chickens inoculated with NRTUAs for 3 days， which aimed to identify differentially expressed genes （DEGs）. Then， the identified DEGs were conducted by GO analysis， enrichment analysis of KEGG signaling pathways， and protein-protein interaction network analysis. The results showed that：1） 964 DEGs were screened， of which 517 genes were upregulated and 447 genes were downregulated. 2） Through GO functional annotation， it was found that differentially expressed genes were significantly enriched in immune response， defense response， response to external stimulus， response to stimulus， extracellular region， cell surface， and other categories. 3） KEGG signaling pathway enrichment analysis revealed that DEGs were mainly involved in pathways such as cytokine-cytokine receptor interactions， intestinal immune network for IgA production， cell adhesion molecules， Toll-like receptor signaling pathway. 4）Through protein-protein interaction network analysis， nine important key molecules were analyzed ， among which IL-6 and IL-18 molecules had the function of regulating cell differentiation and promoting interferon production. IRF1 and IRF7 can regulate the activation of macrophages and other cells as well as antigen presentation ability. SOCS1 can prevent excessive inflammatory response caused by overactive STAT1 signaling pathway. CCL4， CCR5 and CCL20 as chemokines， can induce infiltration of various immune cells in inflammatory areas. CD40 can enhance antigen presentation and regulate the immune function of the body. In conclusion， stimulation of the chicken thymus with NRTUAs at the transcriptional level resulted in a reduction in thymocyte proliferation activity， enhanced activation of T cells， and increased secretion of cytokines. The molecules IL-6， IL-18， IRF1， IRF7， SOCS1， CCL4， CCR5， CCL20 and CD40 played crucial roles in the regulation of NRTUAs in the chicken thymocyte cells. This study provids a theoretical foundation for further exploring the molecular mechanisms of NRTUAs regulating the innate immune response in poultry thymus. |
| Key words: nonreplicating Toxoplasma uracil auxotrophs transcriptomics chickens thymus |
