IL1R1基因敲除对于柯萨奇病毒A10感染C57BL/6乳鼠的影响  

Influence of IL1R1 Knockout on Coxsackievirus-A10 Infected C57BL/6 Mice

作　　者：刘长坤 贺雅娜 张子寒 胡云光[1] 郑惠文[1,3] 刘龙丁 LIU Changkun;HE Yana;ZHANG Zihan;HU Yunguang;ZHENG Huiwen;LIU Longding(Institute of Medical Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Kunming 650118,China;Kunming Medical University,Kunming 650500,China;Key Laboratory of Systemic Innovative Research on Virus Vaccine,Chinese Academy of Medical Sciences,Kunming 650118,China)

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,昆明650118 [2]昆明医科大学,昆明650500 [3]中国医学科学院病毒疫苗研发系统创新研究重点实验室,昆明650118

出　　处：《病毒学报》2024年第2期326-336,共11页Chinese Journal of Virology

基　　金：云南省科技人才与平台计划(项目号:202305AC160008),题目:中青年学术和技术带头人后备人才项目;云南省科技计划-基础研究专项(项目号:202401AS070047),题目:EV-A71和CVA16抗原预存免疫对CV-A10和CV-A6抗原诱导的B细胞和T细胞免疫的影响;云南省科技计划-生物医药专项(项目号:202102AA310043),题目:新型冠状病毒及其疫苗诱导的免疫学特征研究;昆明市春城计划(项目号:2022SCP010),题目:春城科技领军人才项目。

摘　　要：本研究通过柯萨奇病毒A组10型(Coxsackievirus A10, CV-A10)感染C57BL/6野生型乳鼠和白细胞介素-1受体1型(Interleukin 1 Receptor 1,IL-1R1)缺失的C57BL/6乳鼠,比较两者之间的临床症状、病毒体内增殖特征和病理特征差异,以探讨IL-1R1基因缺失对于CV-A10感染乳鼠的影响。本研究采用颅内方式感染C57BL/6野生型和IL-1R1^(-/-)乳鼠,每日记录乳鼠临床症状及死亡情况;两组乳鼠感染后于1、3、5、7、9d处死,并采集各组织样本。通过实时荧光定量PCR(Real-time reverse transcription-quantitative polymerase chain reaction,RT-qPCR)检测组织内病毒基因拷贝数;使用HE染色检测乳鼠组织的病理学变化;RT-qPCR检测病毒感染乳鼠后的不同时间点组织内的细胞因子mRNA转录水平;利用免疫荧光分析对IFN-γ、IL-1β、IL10的细胞因子表达进行分析。结果发现相比于空白对照组,随着感染进程的增加,两组乳鼠均表现出发育迟缓、肢体麻痹等症状;另外在感染后第5d,与野生型乳鼠相比,IL-1R1^(-/-)乳鼠心脏、肺脏和肾脏组织的CV-A10病毒载量均高于野生型乳鼠,且差异具有统计学意义;通过HE染色分析发现,在感染后期IL-1R1^(-/-)乳鼠的肺组织病变严重程度高于同时期野生型乳鼠。在CV-A10感染中后期IL-1R1^(-/-)乳鼠的细胞因子mRNA转录水平明显升高,然而野生型乳鼠的细胞因子mRNA主要在感染后1-2天呈现一过性增加。综上表明,C57BL/6野生型和IL-1R1^(-/-)乳鼠均对CV-A10病毒易感;与野生型乳鼠相比,IL-1R1^(-/-)乳鼠的组织损伤更严重,特别是肺组织的损伤主要发生在CV-A10病毒感染的中晚期;此外,IL-1R1缺陷小鼠在感染后期的各组织器官中检测到持续性的细胞因子表达。This study compared differences in the clinical features,viral proliferative dynamics and pathological characteristics between coxsackievirus(CV)-A10-infected C57BL/6 wild-type suckling mice and CV-A10-infected interleukin 1 receptor 1(IL-1R1)-deficient suckling mice.We investigated the influence of ILIR1 deletion on CV-A10-infected suckling mice.C57BL/6 wild-type and IL-1R1^(-/-)lactating mice were infected intracranially.The clinical symptoms and mortality of newborn mice were recorded daily.The two groups of newborn mice were sacrificed on 1,3,5,7 and 9 days after infection,and samples of various tissues were collected.The viral gene copies of different tissues were detected by real-time reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Staining(hematoxylin and eosin)was used to detect pathologic changes in different tissues of suckling mice.Cytokine transcription levels in tissues were measured by real-time RT-qPCR.Immunofluorescence staining was used to measure expression of the cytokines interferon-γ,interleukin(IL)-1βand IL10.Compared with the blank control group,suckling mice of both groups showed developmental delay and limb paralysis during infection.The viral loads of IL-1R1^(-/-)suckling mice were higher than those of wild-type suckling mice.The difference in viral load between the two groups was significant in heart,lung and kidney tissue 5 days post-infection.Histopathology showed that lesion severity in IL-1R1^(-/-)suckling mice in the late stage of infection was higher than that in wild-type suckling mice at the same time.mRNA transcription of cytokines of IL-1R1^(-/-)newborn mice was increased in the middle and late stages of CVA10 infection,but mRNA transcription of wild-type mice increased transiently 1-2 days after CV-A10 infection.Therefore,C57BL/6 wild type and IL-1R1^(-/-)lactating mice were susceptible to CV-A10.Compared with wild-type mice,inflammatory-tissue injuries of IL-1R1^(-/-)lactating mice occurred mainly in the middle and late stages of CV-A10 infection,and

关 键 词：柯萨奇病毒A10 病毒感染 IL-1R1-/-乳鼠 C57BL/6乳鼠 

分 类 号：R373.23[医药卫生—病原生物学]