利用Aag2细胞过表达体系筛选埃及伊蚊对DENV2易感相关免疫基因

埃及伊蚊Aedes aegypti是登革病毒的主要传播媒介,埃及伊蚊感染登革Ⅱ型病毒(dengue virus typeⅡ,DENV2)后产生先天免疫反应的相关研究,有助于进一步理解埃及伊蚊感染DENV2的过程和机制。本研究通过埃及伊蚊和Aag2细胞感染DENV2进行转录组分析,与未染毒的蚊虫和细胞进行对比,筛选出12个表达变化较大的基因,之后分别构建含有这12个基因的PSL1180polyUBdsRED质粒载体,并使用细胞转染技术在Aag2细胞中建立目标基因的瞬时转染过表达模型,观察了目标基因过表达后细胞上清中DENV2的复制水平。结果显示,12个基因中羟基类固醇脱氢酶样蛋白、蜕皮激素20单加氧酶、3β-羟基类固醇脱氢酶3个基因过表达后细胞上清中病毒拷贝数出现了具有统计学意义的降低,表明这3个基因的表达可能影响DENV2在Aag2细胞中的复制。这3个基因的相关蛋白都涉及到20-羟基脱皮酮的合成,这提示20-羟基脱皮酮可能在蚊虫感染DENV2时介导了重要的免疫调节途径,为进一步研究蚊虫与虫媒病毒的互作机制提供了新的思路。

Mosquito CYP4C21 knockout reduces dengue virus and Zika virus replication in Aedes aegypti cells

Aedes aegypti(Ae.aegypti)is a major vector of dengue virus(DENV)and Zika virus(ZIKV).Understanding the complex interaction mechanisms between mosquito vectors and arboviruses is essential to interrupt virus trans-mission.This study constructed CYP4C21 knockout(KO)Aag2 cells(Ae.aegypti cells)and confirmed that CYP4C21 KO reduced DENV2 and ZIKV copies in Aag2 cells,which suggests that CYP4C21 may play an impor-tant role in mosquito infection with arboviruses.Furthermore,it is the first report of the CYP4 family related to viral infection,which lays the foundation for exploring the role of the CYP4C21 in the interaction of Ae.aegypti and arbovirus and provides novel insights into the function of cytochrome family proteins.