烟曲霉孢子侵染小鼠骨髓源巨噬细胞的转录组研究

Transcriptome study of mouse bone marrow-derived macrophages infected by Aspergillus fumigatus spores

目的揭示小鼠骨髓源巨噬细胞(bone marrow-derived macrophages,BMMs)被烟曲霉(Aspergillus fumigatus)A1160c孢子侵染之后与未处理对照组BMMs相比表达出现差异的免疫信号通路及孢子吞噬和清除等相关基因。方法提取小鼠原代骨髓细胞并将其诱导分化成巨噬细胞。烟曲霉孢子感染并分别提取感染组和对照组BMMs的总RNA,进行RNA测序和分析比较。结果提取小鼠原代骨髓细胞并成功将其诱导分化成BMMs。BMMs在感染烟曲霉孢子后,表面受体Tlr2和C型凝集素受体Clec4e和Clec5a的转录被激活。NF-κB信号传导通路也被激活,如Il1b、Tnf、Nfkb1等炎症因子基因和Jun、Gsk3b、Tec、Cd83等相关基因转录上调。同时,Cxcl1、Cxcl2、Ccl4等多种趋化因子基因转录水平也上调。另外,活性氧产生相关基因Bax、Nos2、Glul、Nlrp3的转录水平明显升高。结论在感染烟曲霉A1160^(c)孢子过程中,BMMs通过上调Toll样受体和C型凝集素受体基因转录水平激活NF-κB等免疫信号传导通路,促进趋化因子及活性氧产生相关基因的表达,引发对烟曲霉孢子的免疫响应和清除。

Objective To reveal the important genes related to phagocytosis,immune responses,and clearance of spores in the mouse bone marrow-derived macrophages(BMMs)infected by Aspergillus fumigatus.Methods Mouse primary bone marrow cells were extracted,induced to differentiate into macrophages,and challenged with A1160^(c) conidia.After infection,the total RNAs of BMMs in the infected group and the control group were extracted and analyzed by RNA-sequencing.Results The primary mouse bone marrow cells were successfully induced to differentiate into BMMs.Compared to untreated control BMMs,the transcription of Toll-like receptor(TLR)Tlr2 and C-type lectin receptors Clec4e and Clec5a were upregulated in conidia-infected BMMs.The NF-κB pathway was also activated through increasing the transcription levels of inflammation related genes such as Il1b,Tnf,Nfkb1,Tnfsf14,Jun,Gsk3b,Tec and Cd40,etc.Meanwhile,the expression of chemokines such as Cxcl2,Cxcl10 and Ccl3 also increased.In addition,the transcription levels of Bax,Nos2,Glul,and Nlrp3,which were related to the generation of reactive oxygen species(ROS),were obviously upregulated.Conclusion To adapt infection of A.fumigatus spores,BMMs activated TLR and C-type lectin receptors,NF-κB pathway,and other immune pathways,exhibiting an elevated immune response and spore clearance potential.