埃及伊蚊唾液源yellow-c蛋白促进登革2型病毒感染的分子机制研究

蚊虫唾液可促进虫媒病毒对宿主感染、致病与传播。然而，单个唾液蛋白的生物学功能知之甚少。Yellow-c（Aael_YPc)是埃及伊蚊唾液源蛋白，前期研究显示吸血、DENV2感染可诱导其表达上调，rAael_YPc 可促进DENV2和ZIKV在宿主细胞内的复制。为了探讨rAael_YPc促进DENV2复制的分子机制，本研究首先采用ELISA、CO-IP、VOPBA等技术观察rAael_YPc与DENV2、巨噬细胞的结合初步探讨病毒的入侵机制；其次采用单细胞测序技术/RNAseq技术构建rAael_YPc作用前后DENV2感染细胞与组织的全转录组差异表达谱，筛选调控宿主注射局部免疫微环境的关键靶分子，进而采用抗体阻断剂/激活剂观察靶分子在rAael_YPc促进病毒复制中的作用；最后采用免疫组化技术和FACS技术初步探讨rAael_YPc作用下，在宿主注射局部DENV2感染细胞种类与组成。

Mosquito saliva enhances arbo-virus infection, immunpathogenesis and transmission. However, little is known on its function for a single saliva protein. Yellow-c (Aael_YPc) is a saliva protein of Aedes aegypti. Previous studies showed the expression of Aael_YPc could be up-regulated by blood-feeding and DENV infection. recombinant Aael_YPc(rAael_YPc) could enhance DENV2 and ZIKV replication in mammal cells. In this study, a combination experiment between rAael_YPc and DENV or rAael_YPc and macrophage will be performed firstly by ELISA, CO-IP, VOPBA; then a differential transcriptome expression profiles of DENV2 infection with or without rAael_YPc will be constructed by single cell sequence or RNAseq. Key molecules of modulating immuno-micro-environment at the inoculated sites will be selected and verified. The mechanism involved in enhancing virus replication of those molecules will be further discussed by antibody blocking and agonists. At last, the types and composition of DENV2 infected cells will be observed by immunohistochemical methods and flow cytometry at the inoculated sites .