IL2rg^(-/-)大鼠支持人RSV在体内的长期感染  

IL2rg^(-/-)rats support prolonged infection of human RSV

作　　者：熊芮 吴勇 杨艳伟[3] 屈哲[3] 刘甦苏[1] 王誉雅 马丽颖[1] 付瑞[1] 彭宜红[2] 梁春南[1] 范昌发[1] XIONG Rui;WU Yong;YANG Yanwei;QU Zhe;LIU Susu;WANG Yuya;MA Liying;FU Rui;PENG Yihong;LIANG Chunnan;FAN Changfa(Institute for Laboratory Animal Resources,National Institutes for Food and Drug Control,National Rodent Laboratory Animal Resources Center,Beijing 102629,China;Department of Microbiology&Infectious Disease Center,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100083,China;National Center for Safety Evaluation of Drugs,Institute for Food and Drug Safety Evaluation,National Institutes for Food and Drug Control,Beijing 100176,China)

机构地区：[1]中国食品药品检定研究院实验动物资源研究所(国家啮齿类实验动物资源库),北京102629 [2]北京大学医学部基础医学院病原生物学系,北京100083 [3]中国食品药品检定研究院安全评价研究所(国家药物安全评价监测中心),北京100176

出　　处：《中国实验动物学报》2024年第1期17-24,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家重点研发计划(2021YFC2301700);国家科技重大专项(2017ZX10304402)。

摘　　要：目的为了克服已有人呼吸道合胞病毒(human respiratory syncytial virus,hRSV)动物模型的局限性,如半受纳性和感染持续时间短,本文利用TALEN基因编辑技术建立了IL2rg基因敲除(IL2rg^(-/-))的大鼠模型。方法用hRSV滴鼻感染该动物模型,观察感染期(0~35 d)的临床表征、体重及体温变化;记录不同时间点(滴鼻感染后第4、11、20、35天)鼻腔、气管、肺等呼吸道脏器的病毒总拷贝数;在观察终点(滴鼻感染后第35天)对感染动物的靶器官进行病理分析;观察不同时间点(滴鼻感染后第4、20、35天)外周血T、B、NK、NKT细胞的变化及不同时间点多种细胞因子的变化。结果(1)通过鼻内接种hRSV后,纯合的IL2rg基因敲除大鼠的呼吸道内能保持较高的病毒载量,鼻腔中病毒的平均峰值滴度能快速升至1×10^(10 )copies/g,至第5周时,病毒依然能维持复制,病毒载量亦可达到1×10^(7)copies/g。(2)但其鼻、气管和肺组织,无明显病变。(3)感染hRSV的IL2rg^(-/-)大鼠外周血B细胞含量有上升。(4)IL-6和MCP-1两种细胞因子都在感染前期上升,在观察终点回落。结论本研究基于TALEN技术建立了IL2rg^(-/-)大鼠模型,并发现该模型能很好地支持hRSV高水平复制和并长期感染,为抗病毒药物筛选、抗hRSV抗体体内效力评价,提供了良好的动物模型。Objective To overcome the limitations of existing human respiratory syncytial virus(hRSV)animal models,such as semi-permissiveness and short duration of infection,this study established an IL2rg gene knockout IL2rg^(-/-))rat model using TALEN gene editing technology.Methods The animal model was infected with hRSV intranasally.Clinical characteristics,body weight,and temperature changes were observed over the infection period(0~35 days).The total viral loads in respiratory organs,such as the nasal tissue,trachea,and lungs,were measured at various time points(4,11,20,and 35 days post-infection).Pathological analysis was conducted on target organs at the endpoint of observation(35 days post-infection).Changes in peripheral blood T,B,NK,and NKT cells and various cytokines were assessed at various time points(4,20,and 35 days post-infection).Results(1)IL2rg^(-/-)knockout rats sustained high viral loads in the nasal cavity upon intranasal inoculation with hRSV.The average peak titer rapidly reached 1×10^(10) copies/g in nasal tissue and 1×10^(7) copies/g up to 5 weeks post-infection.(2)However,no significant pathological changes were noted in nasal,tracheal,or lung tissues.(3)An increase was observed in the content of peripheral blood B cells in hRSV-infected IL2rg^(-/-)rats.(4)IL-6 and MCP-1 were increased in the early stage of infection and then decreased at the end of the observation period.Conclusions This study established a new IL2rg^(-/-)rat model using TALEN technology and found that this model effectively supported high-level replication and long-term infection of hRSV,providing a good basis for antiviral drug screening and in vivo efficacy evaluation of anti-hRSV antibodies.

关 键 词：IL2rg-/-大鼠 TALEN基因编辑技术 NK细胞缺陷 人呼吸道合胞病毒 感染动物模型 

分 类 号：Q95-33[生物学—动物学]