PRRSV對豬肺泡巨噬細胞天然免疫功能影響的分子機制.pdf

1、Y773592分類號：密級；’j目巖≮式普博士學位論文單位代碼；10019學號：B02212PRRSV對豬肺泡巨噬細胞天然免疫功能影響的分子機制MolecularMechanismoftheEffectofPorcineReproductiveandRespiratorySyndromeVirusonInnateImmunityofPorcinePulmonaryAlveolarMacrophages研究生：羞登指導教師：塹絲盎基燕合作

2、指導教師：申請學位門類級別：盤堂壤專業(yè)名稱：亟墮薹匡莖壹些研究方向：墨墊僉i癌圭塋墨盤塞鱟所在學院：麴塹堡鱟睦2005年5月AbstractPorcinereproductiveandrespiratorysyndrome，whichischaracterizedbyrespiratoryreproductivedisordersofsowandrespiratorydiseaseofpiglets，isaviraldiseasecau

3、sedbyporcinereproductiveandrespiratorysyndromevirus(PRRSV)InthisstudytheeffectofPRRSVinfectiononimmunityofporcinepulmonaryalveolarmacrophages(PAM)wassystematicallyinvestigatedfromtheviewofinnateimmunityTheresearchinvolve

4、dinanalysisofthemRNAtranscriptionofpattemrecognitionreceptor(PRR)moleculesassociatedwithinnateimmunityIFNstimulatedgenes(ISG)andcytokinesinPAMinfectedwithPRRSVTheaimofourstudyistoexplorethemolecularmechanismoftheeffectof

5、PRRSVinfectiononPAMimmunityPartialgenefragmentsofswinepRRincludingCDl6，CDl8，TLR2，TLR4，SR，moleculesassociatedwithinnateimmunityincludingNrampl，SPA，ISGincludingOAS，PKR，Mxl，andcytokinesinvolvedIFNⅡ，1FN7，IL113，IL一6，TNF—n，IL8

6、，MCP1，IL一10，lL12p40，IL一18andGMCSFwereamplifiedbyRTPCRfromPAMandclonedaccordingtotheirsequencesavailableinGenBankrespectivelyAftersequencing，competitivedeletedclonesfortheeDNAmoleculesmentionedaboveweregeneratedbyrcverseP

7、CRThenthelinearregressionequationswereobtainedafterconstructionofthestandardcompetitivecurvesrespectivelybyquantitativecompetitivePCR(qcPCR)assayThePAMfromPRRSVinfectedPAMandcontrolPAMinvitroandinvivowereinvestigatedfort

8、hemRNAtranscriptionsofPRRandmoleculesassociatedwithhm砒eimmunitybyqcPCRaccordingtotheirrespectivestandardcompetitivecorveconstructed／nvitro，allPRRandmoleculesassociatedwithinnateimmunityincludingCDl6，CDt8，TLR2，TLR4，SR，Nra

9、mpl，SPAdecreasedsharplytothelowestlevelat4hpostinoculation(P1)，thenincreasedgraduallytonormalat48hPI／nvivo，allmoleculesdecreasedatdifferentlevelduringtheearlystageofPRRSVinfection(atday314PI)Inconclusion，themRNAtranscrip

10、tionsofthesemoleculeswereprohibitedduringtheearlyinfection，resultingintheimpairmentofmacrophageinnateimmunityDuringthelatestageofPRRSVinfection(atday28—42PI)，themRNAtranscriptionsofthesemoleculesrecoveredtonormalleveloru

11、pregulated，suggestingthefunctionofPAMappearsteboundandincreasesomewhatThemRNAtranscriptionsof1SGofPRRSVinfectedPAMandcontrolPAM加vitroandinvivowereinvestigatedbyqcPCRaccordingtotheirrespectivestandardcompetRivecurveconstr

12、ucted／nvitro，allISGmRNAtranscriptionsincludingOAS，PKRandMxlinPRRSVinfectedPAMshowedlowerlevelthanthatinthecontrolPAM，exhibitingthatthemRNAtranscriptionsofthesegenesweresuppressedbyPRRSVinfectionThishintsthatPRRSVinfectio

13、nmightdamagetheantiviralfunctionofPAM／nvivoISGmRNAtranscriptionsinPAMofPRRSVinfectedpigletshadnosignificantchangecomparedtothatofcontrolpigletsatday3and7PI；howevertheyshowedasharpdecreaseatday14PI，andthenascendedtonormal