创伤弧菌致死性感染小鼠模型的组织病理改变及其在疫苗中的应用初探  

Pathological response of a mouse model of lethal Vibrio vulnificus infection and its preliminary application in inactivated whole cell vaccine

作　　者：朱宝行 潘佳乐 刘姝琳 叶演 宋振 李雨贤 杨赟 孙红武[1] 邹全明[1] 彭六生[1] ZHU Baohang;PAN Jiale;LIU Shulin;YE Yan;SONG Zhen;LI Yuxian;YANG Yun;SUN Hongwu;ZOU Quanming;PENG Liusheng(Department of Microbiology and Biochemical Pharmacy,Faculty of Pharmacy and Laboratory Medicine,Army Medical University(Third Military Medical University),Chongqing;College of Pharmacy,Chongqing University of Technology,Chongqing,China)

机构地区：[1]陆军军医大学(第三军医大学)药学与检验医学系微生物与生化药学教研室,重庆 [2]重庆理工大学药学院,重庆

出　　处：《陆军军医大学学报》2025年第7期656-663,共8页Journal of Army Medical University

基　　金：国家重点研发计划(2021YFC2302603)。

摘　　要：目的建立创伤弧菌最低致死剂量感染小鼠模型,评估创伤弧菌灭活全菌疫苗的免疫保护效果。方法采用不同剂量的创伤弧菌腹腔注射小鼠,确定感染的最低致死剂量;采用组织匀浆涂板检测脏器的细菌定植量,组织切片染色分析脏器的病理改变;将肝脏消化成单细胞悬液,采用流式细胞术检测免疫细胞的应答变化;制备创伤弧菌灭活全菌疫苗,采用不同途径免疫小鼠,观察疫苗的保护效果。结果成功确定了创伤弧菌腹腔感染的最低致死剂量为1×10^(6)CFU,创伤弧菌感染后主要定植在小鼠的肝脏,并造成了肝脏的严重病理损伤;与未感染组小鼠相比,感染组小鼠肝脏中的中性粒细胞比例显著增加,而B细胞和T细胞的比例则相应的显著下降(P<0.05);小鼠经肌肉或腹腔单次注射灭活全菌疫苗7 d后即可有效抵抗创伤弧菌的致死性感染(P<0.01),但血清IgG抗体的水平并未明显升高。结论成功确定了创伤弧菌最低致死剂量感染小鼠模型及其组织病理学应答特征,灭活全菌疫苗可能通过IgG抗体非依赖的方式介导对该创伤弧菌感染小鼠的快速免疫保护。Objective To establish a mouse model of infection with the minimum lethal dose of Vibrio vulnificus(V.vulnificus)and to evaluate the protective efficacy of inactivated whole-cell(IWC)vaccine using this model.Methods A mouse model of lethal-dose infection was established by intraperitoneal injection of different doses of V.vulnificus.Bacterial colonization in the organs was detected with tissue homogenate plating,and pathological changes in the organs were observed after tissue section staining.Flow cytometry was used to detect immune cell responses after liver tissues were digested into single-cell suspension.IWC vaccine of V.vulnificus was prepared,and the mice were immunized through different routes to observe the protective efficacy of the vaccine.Results A mouse model of infection with the minimum lethal dose at 1×10^(6)CFU of V.vulnificus was successfully established.After infection,the bacteria were mainly colonized in the liver of mice and caused severe pathological damages.Compared with the uninfected mice,the proportion of neutrophils in the liver was significantly increased in the infected mice,whereas the proportions of B cells and T cells were correspondingly decreased(P<0.05).A single intramuscular or intraperitoneal injection of the IWC vaccine could protect the mice effectively against lethal infection of V.vulnificus in 7 d later(P<0.01),although the level of serum IgG having no significant increase.Conclusion A mouse model of lethal-dose infection with V.vulnificus is successfully established,with histopathological characteristics.The IWC vaccine of V.vulnificus rapidly mediates immune protection in this model probably independent of IgG.

关 键 词：创伤弧菌 小鼠模型 肝损伤 灭活全菌疫苗 

分 类 号：R-332[医药卫生] R378.3R392.7