Song Li, Xiong Dan, Wen Yaya, Tan Ruimeng, Kang Xilong, Jiao Xinan, Pan Zhiming
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China.
Curr Issues Mol Biol. 2023 Mar 27;45(4):2798-2816. doi: 10.3390/cimb45040183.
The flagellin (FliC) of is a potential vaccine adjuvant as it can activate innate immunity and promote acquired immune responses. Macrophages are an important component of the innate immune system. The mechanism of flagellin's adjuvant activity has been shown to be related to its ability to activate macrophages. However, few studies have comprehensively investigated the effects of flagellin in macrophages using transcriptome sequencing. In this study, RNA-Seq was used to analyze the expression patterns of RAW264.7 macrophages induced by FliC to identify novel transcriptomic signatures in macrophages. A total of 2204 differentially expressed genes were found in the FliC-treated group compared with the control. Gene ontology and KEGG pathway analyses identified the top significantly regulated functional classification and canonical pathways, which were mainly related to immune responses and regulation. Inflammatory cytokines (IL-6, IL-1β, TNF-α, etc.) and chemokines (CXCL2, CXCL10, CCL2, etc.) were highly expressed in RAW264.7 cells following stimulation. Notably, flagellin significantly increased the expression of interferon (IFN)-β. In addition, previously unidentified IFN regulatory factors (IRFs) and IFN-stimulated genes (ISGs) were also significantly upregulated. The results of RNA-Seq were verified, and furthermore, we demonstrated that flagellin increased the expression of IFN-β and IFN-related genes (IRFs and ISGs) in bone marrow-derived dendritic cells and macrophages. These results suggested that flagellin can activate IFN-β-related immune responses in macrophages, which provides new insight into the immune mechanisms of flagellin adjuvant.
[细菌名称]的鞭毛蛋白（FliC）是一种潜在的疫苗佐剂，因为它可以激活先天免疫并促进获得性免疫反应。巨噬细胞是先天免疫系统的重要组成部分。鞭毛蛋白的佐剂活性机制已被证明与其激活巨噬细胞的能力有关。然而，很少有研究使用转录组测序全面研究[细菌名称]鞭毛蛋白对巨噬细胞的影响。在本研究中，RNA测序用于分析FliC诱导的RAW264.7巨噬细胞的表达模式，以确定巨噬细胞中的新转录组特征。与对照组相比，FliC处理组共发现2204个差异表达基因。基因本体论和KEGG通路分析确定了最显著调节的功能分类和经典通路，这些主要与免疫反应和调节有关。刺激后，炎症细胞因子（IL-6、IL-1β、TNF-α等）和趋化因子（CXCL2、CXCL10、CCL2等）在RAW264.7细胞中高表达。值得注意的是，鞭毛蛋白显著增加了干扰素（IFN）-β的表达。此外，以前未鉴定的干扰素调节因子（IRF）和干扰素刺激基因（ISG）也显著上调。RNA测序结果得到验证，此外，我们证明鞭毛蛋白增加了骨髓来源的树突状细胞和巨噬细胞中IFN-β以及IFN相关基因（IRF和ISG）的表达。这些结果表明，[细菌名称]鞭毛蛋白可以激活巨噬细胞中与IFN-β相关的免疫反应，这为鞭毛蛋白佐剂的免疫机制提供了新的见解。
