家蝇Eiger/Wengen信号的免疫调控机制

在脊椎动物中，肿瘤坏死因子/肿瘤坏死因子受体(TNF/TNFR)系统参与多种生理病理过程，例如肿瘤、炎症和免疫等。Eiger/Wengen是最早从果蝇体内发现的唯一一对TNF/TNFR同源蛋白。Eiger突变会影响果蝇的免疫反应能力，但具体机制尚不清楚。本课题组通过转录组测序和基因数字表达谱分析，筛选到多个家蝇TNF通路成员基因，并已证实它们参与了家蝇免疫调控。本项目拟首先分析Eiger、Wengen和TRAF在家蝇体内的时空分布及亚细胞定位，以了解它们的表达模式。然后利用RNAi和CRISPR/Cas9技术对幼虫体内Eiger等基因进行敲低或敲除，观察抗菌肽表达、黑化反应和细胞吞噬作用的变化。利用重组Eiger和共转染技术在S2细胞内研究Eiger、Wengen和TRAF等对抗菌肽表达的影响，探讨Eiger/Wengen信号对免疫反应的调控模式以及与Toll和Imd通路间的联系。

The tumor necrosis factor/receptor (TNF/TNFR) system participates in diverse physiological and pathological events in vertebrates, such as cancer, inflammation and immune responses. Eiger and Wengen are found to be the sole counterparts of the TNF/TNFR superfamily members in Drosophila. Loss-of-function eiger mutants of Drosophila show lower immunity, but the underlying mechanism remains unknown. Base on the transcriptome and digital gene expression information, we identified a few members of the TNF pathway in Musca domestica. Preliminary test results imply these genes take part in the immune regulation of housefly. To understand the function of Eiger/Wengen pathway in immune system, the express profiles of Eiger, Wengen and TRAF will be investigated by Western blot, ISH, subcellular localization and Real-time PCR. The changes of AMP expression, melanization and phagocytosis will be detected following knowckout or knockdown of Eiger, Wengen and TRAF in housefly by RNAi and CRISPR/Cas9. Recombinant Eiger and cotransfection technology will be employed to analysis the affection of Eiger, Wengen and TRAF on the expression of AMPs in S2 cell. The main objective is to understand the regulation of Eiger/Wengen pathway on immune system and the relation between Eiger/Wengen pathway and Toll or Imd pathway.