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1、萬方數(shù)據(jù)生堡塞墅塑墮盎題垂堂塑查!Q!!生!旦筮望鲞筮!塑墾!!!!望!墾翌墾!!!!!竺!:也竺!!!!:!!!:!!:墮!:!流感病毒H3N2小鼠適應(yīng)株的建立及分子機(jī)制探究胡云光,徐娟,胡雅潔,宋杰,乞素冬,秦曉峰,劉龍丁【摘要】目的建立季節(jié)性甲型流感病毒influenzaAvirus[(A/HongKong/1968(H3N2)]毒株的鼠肺適應(yīng)株,并對其適應(yīng)過程的分子機(jī)理進(jìn)行初步探究。方法以滴鼻的方式使小鼠感染H3N2流感病毒,并在小鼠肺部進(jìn)行連續(xù)傳代,通過對小鼠的臨床觀察、病毒載量及病毒滴度測定、肺部病理改變、細(xì)胞因子
2、檢測等方面的觀察和檢測,來獲得季節(jié)性流感病毒influenzaAvirus[A/HongKong/1968(H3N2)]毒株的鼠肺適應(yīng)株。結(jié)果人季節(jié)性甲型流感病毒influenzaAvirus[A/HongKong/1968(H3N2)]毒株,經(jīng)過小鼠肺中連續(xù)7次傳代以后,病毒毒力逐漸增強(qiáng),鼠肺中病毒載量提高了4個(gè)數(shù)量級,病毒滴度上升了2.519CCID。對流感病毒HA、NA兩個(gè)節(jié)段的測序,發(fā)現(xiàn)這兩個(gè)節(jié)段上均有一個(gè)有義突變。結(jié)論在自然狀態(tài)下并不感染小鼠的野生季節(jié)性流感病毒H3N2毒株可以通過在小鼠肺部中連續(xù)傳代,得到毒力較高的
3、鼠肺適應(yīng)株,HA節(jié)段中248位的Ile突變?yōu)閂al,NA節(jié)段中444位的Val突變?yōu)镮le,這兩個(gè)突變位點(diǎn)可能與流感病毒毒力相關(guān)?!局黝}詞】流感病毒A型;小鼠,近交BALBC;病毒培養(yǎng)Establishmentofamouse-adaptedseasonalinfluenzaAH3N2virusstrainandpreliminaryinquiryitsmolecularmechanismHuYunguang,XuJuan,HuYajie,SongJie,QiSudong,QinXiaofeng,LiuLongding.Chi
4、neseAcademyofMedicalSciencesPekingUnionMedicalCollegelnsituteofMedicalBiology,Kunming650118,ChinaCorrespondingauthor:QinXiaofeng,Email:fxfqin@gmail.corn;LiuLongding,Email:longdingl@gmail.corn【Abstract】ObjectiveToestablishadaptedstrainofseasonalinfluenzavirusinfluenz
5、aAvirus[A/HongKong/1968(H3N2)]inmiceandexplorethemolecularmechanismofadaptionpreliminary.MethodsThemicemodelwasinducedbynosedroppingwithwildtypeH3N2influenzavirus,bycontinuouspassageinthelungofmiceanddetectionoftheclinicalmanifestation,viralload,virustiter,pathology
6、,cytokines,togettheadaptedstrainofseasonalinfluenzaAvirus[A/HongKong/1968(H3N2)].ResultsAfter7timescontinuouspassageinthelungofmice,virulencewasincreased,viralloadandvirustiterwereincreasedeither.GenomesequencingandalignmentindicatedthattheHAandNAgenewasmutated.Conc
7、lusionTheinfluenzavirusH3N2ofmicelungadaptioncanbeacquiredfromwildtypeseasonalH3N2bycontinuouspassageinthelungtissuesofmice.MutationofIletoValatresidue248ofHAandValtoIleatresidue4440fNAproteinmaybethekeyvirulencedeterminant.【Keywords】InfluenzaAvirus;Mice,inbreadBALB
8、C;VircscuhivastionDOI:10.3760/cma.j.issn.1003-9279.2015.03.004基金項(xiàng)目:國家自然科學(xué)基金面上項(xiàng)目(81373142);云南省生物重大科技專項(xiàng)(2012ZA009)作者單位:中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院醫(yī)學(xué)生物學(xué)研究