鸭坦布苏病毒突破血-脾屏障引发脾免疫微环境变化的机理研究

鸭坦布苏病毒（DTMUV）是一种严重影响我国养鸭业的新发传染病病原。课题组前期实验结果显示，脾脏是DTMUV感染时病变最明显、病毒含量最高的器官。以此为切入点，申请者拟综合运用形态学和分子生物学方法，探究DTMUV突破鸭血-脾屏障（BSB）的动态机制；禽类BSB的高内皮血管是淋巴细胞归巢的发生部位，通过RNA-seq测序技术筛选鉴定不同感染期脾脏淋巴细胞归巢相关差异表达基因，同时利用荧光双标和激光共聚焦技术厘清淋巴细胞与高内皮归巢分子的互作关系，进而分析DTMUV感染对脾脏淋巴细胞归巢过程的影响。在此基础上，通过显示脾脏各种免疫细胞分布和细胞因子表达变化，进一步揭示DTMUV感染条件下鸭脾结构区域及其免疫微环境改变的内在规律。研究结果将从新发现的禽类BSB的特殊角度，在组织、细胞和分子水平上阐明DTMUV的感染机制，为制定有效的防控措施和研制特异的DTMUV疫苗提供科学的理论支撑。

Duck Tembusu virus （DTMUV） is a new infectious virous which seriously affects duck industry. Our previous works showed that the spleen was the most diseased organ which was the highest viral load. According to the previours research work, we are going to detect the molecular cell mechanism of DTMUV invasion into duck spleen through the newly reported blood-spleen barrier (BSB). High endothelial blood vessel within the BSB is the location where lymphocyte homing takes place in the poultry, and differentially expressed genes associated to lymphocyte homing are screened and identified by RNA-seq sequencing. Interactions between lymphocytes and High endothelial cells are investigated by using fluorescent double labeling technique and laser screen confocal microscopy. The influences on lymphocyte homing dynamic by DTMUV incursion are further analysed. The inherent law of variation in regional constructions and immune microenvironments of duck spleen is examined under DTMUV infection conditions. The Pathological mechanism of DTMUV infection is going to be elucidated on tissue, cell and molecular levels. All the results will provide the theoretical principle for DTMUV disease prevention and vaccine development.