甲型H1N1流感病毒感染小鼠肺共信号分子变化研究  

Changes of pulmonary co-signaling molecules in influenza A(H1N1)virus infected mice

作　　者：黄锐[1,2] 黄霞 车霄 王乐霄 李佳颖 王志杰 高蓉 王福生 焦艳梅 白冰珂 徐哲 HUANG Rui;HUANG Xia;CHE Xiao;WANG Lexiao;LI Jiaying;WANG Zhijie;GAO Rong;WANG Fusheng;JIAO Yanmei;BAI Bingke;XU Zhe(The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510515,Guangdong Province,China;Senior Department of Infectious Diseases,the Fifth Medical Center,Chinese PLA General Hospital,Beijing 100039,China;Graduate School,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]南方医科大学第二临床医学院,广东广州510515 [2]解放军总医院第五医学中心感染病医学部,北京100039 [3]解放军总医院研究生院,北京100853

出　　处：《解放军医学院学报》2024年第4期405-410,共6页Academic Journal of Chinese PLA Medical School

基　　金：国家重点研发计划(2017YFC1200802)。

摘　　要：背景甲型流感病毒传染性强,重症感染病死率高。共信号分子是参与免疫应答的重要分子。探究甲型流感病毒感染后肺病毒载量与共信号分子的变化特点及相关性可为病毒感染后的治疗提供参考依据。目的探究甲型流感病毒A/Puerto Rico/8/34(H1N1)(PR8株)感染小鼠肺病毒载量和共信号分子的变化特点及相关性。方法72只7周龄BALB/c小鼠随机分为高剂量病毒组(108 TCID50/mL,50μL/只,32只)、低剂量病毒组(104 TCID50/mL,50μL/只,32只)和对照组(PBS,50μL/只,8只),三组分别于感染后24 h、48 h、72 h和96 h每个时间点每组各处死8只、8只、2只小鼠;解剖取左肺行HE染色,实时定量逆转录PCR(qRT-PCR)检测肺病毒载量以及共信号分子CD28、CD226、ICOS、CTLA-4、TIGIT、PD-1 mRNA相对表达量。将高、低剂量病毒组所有病毒感染小鼠(n=64)肺病毒载量与共信号分子mRNA相对表达量行相关性分析。结果感染后96 h内,高剂量病毒组相比低剂量病毒组小鼠肺损伤较重,且共刺激分子CD28、CD226、ICOS mRNA相对表达量均减少(P均<0.05)。高剂量病毒组小鼠感染后24 h PD-1 mRNA相对表达量增加(P<0.01),感染后48 h CTLA-4 mRNA相对表达量增加(P<0.01)且病毒复制达到峰值。低剂量病毒组小鼠感染后72 h PD-1 mRNA相对表达量增加(P<0.05)且病毒复制达到峰值,感染后96 h CTLA-4 mRNA相对表达量增加(P<0.01)。病毒感染小鼠(n=64)肺病毒载量与PD-1 mRNA相对表达量存在正相关关系(r=0.540,P<0.001),与CD226 mRNA相对表达量存在负相关关系(r=-0.340,P=0.006)。结论甲型流感病毒感染小鼠肺组织中CTLA-4、PD-1 mRNA相对表达量增加等现象的早期出现可能提示小鼠感染的重症化。肺组织中的PD-1和CD226可能是与肺病毒载量相关的共信号分子靶点。Background Influenza A virus is highly infectious with a high mortality rate in severe infection cases.Co-signaling molecules are important molecules involved in immune response.Exploring the changing characteristics and correlation between pulmonary viral load and co-signaling molecules after influenza infection can provide reference for the treatment of influenza A virus infection.Objective To explore the changing characteristics of pulmonary viral load and its association with co-signaling molecules mRNA expression in mice infected with influenza A virus A/Puerto Rico/8/34(H1N1)(PR8 strain).Methods Totally 72 BALB/c mice aged 7 weeks were randomly divided into high dose virus group(108TCID50/mL,50μL/mouse,n=32),low dose virus group(104TCID50/mL,50uL/mouse,n=32)and control group(PBS,50uL/mouse,n=8),8,8 and 2 mice were sacrificed in the three groups at each time point of 24 h,48 h,72 h and 96 h after infection,respectively.All mice were dissected for the left lung which used for HE staining,qRT-PCR was applied to detect the pulmonary virus load and the relative mRNA expression levels of cosignaling molecules CD28,CD226,ICOS,CTLA-4,TIGIT and PD-1.The correlation between pulmonary viral load and mRNA expression of co-signaling molecules were analyzed in all mice infected with high and low dose viruses.Results Within 96 h after infection,the lung injury of high dose virus group was more severe than that of low dose virus group,and the mRNA expression of co-stimulatory molecules CD28,CD226 and ICOS was decreased(all P<0.05).In the high dose virus group,the relative expression of PD-1 mRNA increased at 24 h after infection(P<0.01),the relative expression of CTLA-4 mRNA increased(P<0.01)and the viral replication reached to the peak at 48 h after infection.In the low dose virus group,the relative expression of PD-1 mRNA increased(P<0.05)and the viral replication reached to the peak at 72 h after infection,the relative expression of CTLA-4 mRNA increased(P<0.01)at 96 h after infection.The pulmonary viral load of infect

关 键 词：甲型流感病毒 病毒性肺炎 共信号分子 共刺激分子 共抑制分子 

分 类 号：R511.7[医药卫生—内科学]