外文翻譯--麥冬多糖脂質(zhì)體可以提高雞的非特異性和特異性免疫應答（英文）

1、CarbohydratePolymers119(2015)219–227ContentslistsavailableatScienceDirectCarbohydratePolymersjournalhome page:www.elsevier.com/locate/carbpolOphiopogonpolysaccharideliposomecanenhancethenon-specific andspecificimmuneres

2、ponseinchickensYunpengFan a,1, XiaMa b,1, JingZhang c, LinMa a, YuanyuanGao a, WeiminZhang a, XiaopingSong a,?, WeifengHou a, ChaoGuo a, DewenTong a,?a CollegeofVeterinaryMedicine,NorthwestA OPL at highandmediu

3、mdoses could significantlyimprovethe phagocytosicindex, promotelymphocyteprolifera-tion,increasethe proportionof T lymphocytesubpopulations(CD4+ and CD8+), enhanceantibodytiterandimprovethe protectiverate in v

4、ivo.Moreover,its efficacy was significantlybetter thanophiopogonpolysaccharide(OP). Theseresultsindicatedthat the immune-enhancingactivityof OP wassignificantlyimprovedafter encapsulatedwith liposome.©201

5、4 ElsevierLtd. All rightsreserved.1.IntroductionAnimalepidemicshaveseriousandextensiveimpactonbreed-ingindustryinourcountry,whichforinstance,Newcastledisease,Avianinfluenza,porcinereproductiveandrespiratorysyndromeande

6、tc.oftenhappenandbringthebusinessagreatloss.Thesediseasesmostbelongtoviralinfectiousdiseases.Nowadays,vac-cinationisstilltheprincipalmeansforthepreventionandcontrolofviralinfectiousdiseasesowingtothelackofspecifictreatme

7、nt.However,mostvaccinesespeciallythegeneticallyengineeringvac-cinecangetsatisfactoryeffectwiththehelpofimmunopotentiator,whichplayanessentialroleinimprovingtheeffectofvaccine(ZhaoAbbreviations:OP,ophiopogonpolysaccharide

8、;OPL,ophiopogonpolysaccha-rideliposome;ND,Newcastledisease;IL,interleukin;PHA,phytohemagglutinin;MTT,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide;PBS,phos-phatebufferedsaline;DMSO,dimethylsulfoxide;NDV,New

9、castlediseasevirus;VC,vaccinecontrol;BC,blankcontrol;BL,blankliposome;CC,cellcontrol. ? Correspondingauthorsat:CollegeofVeterinaryMedicine,NorthwestAfax:+00862987091032.E-mailaddress:dwtong@nwsuaf.edu.cn(D.Tong).1 Thesea

10、uthorscontributedequallytothiswork.etal.,2014).Whilethechemicalimmunopotentiatorweareusinginclinicpresentlyhavesomedisadvantagessuchaseasytocumu-late,singletarget,greatsideeffectsandsoon(Harandi,Medaglini,Shattock,Ma,Guo

11、,Wang,Hu,Zhangetal.,2013).Asanaturalmacromoleculeactivematerial,polysaccharidecannotonlyactivatetheimmunecells,improvethelevelofantibodies,promotethereleaseofcytokinesandactivatecomplement,butalsoplayimportantrolesinviru

12、scontrolandcancertherapy(Schepetkinetal.,2013;Sheu,Lyu,Lee,&Cheng,2013).Inaddition,polysaccharidewillnotproducesignif-icantsideeffectswhenitregulatestheimmunefunctionofbody,itistheidealimmunopotentiatorinclinicalappl

13、ication.Ophiopogonisjaponicus(RadixOphiopogonisJaponici),asatra-ditionalChinesemedicine,itsusecouldbedatedbackmorethan2000yearsagoandberecordedinShenNong’sMateriaMedica.http://dx.doi.org/10.1016/j.carbpol.2014.11.0480144

14、-8617/©2014ElsevierLtd.Allrightsreserved.Y.Fanetal./CarbohydratePolymers119(2015)219–2272212.4.2.ActivityofOPLonphagocytosisofmacrophagesinvitroThephagocytosisofmacrophageswasmeasuredbyneutralreduptake(Chen,Zhang,Sh

15、en,&Wang,2010).AfterculturedwithOPLorOPfor48h,100?Lofneutralredsolution(0.075%,w/w)wasaddedandincubatedfor4h.Afterdiscardingsupernatant,thecellswerewashedwithPBStwicetoremovetheneutralredwhichwasn’tphagocytizedbyMacr

16、ophages.ThenDMSO(100?Lwell?1)wasaddedtolyseMacrophages.Theabsorbanceat490nmwasassayedbyamicroplatereader.2.4.3.DeterminationofIL-2andIL-6levelsThemacrophageswereinoculatedinto96-wellcultureplates.Afterbeingtreatedwithorw

17、ithoutdrugsfor48h,theplateswerecentrifugedat1000×gfor10min,andthesupernatantwascol-lectedfordeterminingthecontentsofIL-2andIL-6byELISAkit(BiosamiteBiotechnologyCo.Ltd.Shanghai,China),respectively,accordingtothemanuf

18、acturer’sinstructions.2.4.4.ActivityofOPLonphagocyticindexinvivoThephagocyticindexofthemacrophageswasmeasuredbycar-bonclearanceaccordingtotheprocedureofpreviousdescription(Wang,Wang,Li,Shi,&Zhong,2009).Ninety14-day-o

19、ldchickenswererandomlyassignedintosixgroups.Thechickensinfiveexper-imentalgroupswereintramuscularlyinjectedwith1.0mL ofOPLathigh(4.0mg mL?1),medium(2.0mg mL?1)andlow(1.0mg mL?1)dose,OP(4.0mg mL?1)andBL,respectively,

20、inBCgroup,withphys-iologicalsaline,onceadayfor3successivedays.Ondays7and14afterthelastdrugadministration,sixchickenswerechosenran-domlyfromeachgroupandintravenouslyinjectedwithdilutedIndiainkat1mL/100gbodyweight.Bloodsam

21、pleswerecollectedat2min(T2)and10min(T10)fromthebrachialvein,andthen20?Lofbloodwasmixedwith2mL 0.1%Na2CO3.Theabsorbanceat600nmwasmeasuredonaUV–visiblespectrophotometer.Theliverandthespleenwereweighed,andthephagocyticinde

22、xwascalculatedasfollows:K=(lgOD2 ?lgOD10)/(T10 ?T2).Phagocyticindex=K1/3 ×Mbodyweight/(Mliverweight +Mspleenweight).2.5.ActivityofOPLonspecificimmuneresponseThreehundred14-day-oldchickenswererandomlydividedintoseven

23、groupsandvaccinatedwithNDvaccine(LaSotastrain,no.130920,Bio-drugCompanyofVeterinary,BeijingCity)exceptBCgroup,repeatedvaccinationat28-day-old.Atthesametimeofthefirstvaccination,thechickensinfiveexperimentalgroupswereintr

24、amuscularlyinjectedwith1.0mL ofOPLathigh(4.0mg mL?1),medium(2.0mg mL?1)andlow(1.0mg mL?1)dose,OP(4.0mgmL?1)andBL,respectively,invaccinecontrol(VC)andBCgroups,1.0mL ofphysiologicalsaline.Ondays7,14,21,28and35afterthe

25、firstvaccination,thebloodsamplesofsixchickenswerecollectedrandomlyfromeachgroupformeasuringperiph-erallymphocyteproliferationbyMTTmethod,CD4+ andCD8+ Tcellsdistributionbyflowcytometry,andserumhemagglutina-tioninhibition(

26、HI)antibodytiterbymicro-method.Onday35afterthefirstvaccination,thechickensexceptforBCgroupwerechallengedwith0.5mL ofNDV(F48E9strain,suppliedbyChinainstituteofveterinarydrugcontrol)with10LD50 byintramuscularinjection.The

27、pathogenicanddeadstatusesofchickenswereexam-ineddailyfor15successivedaysafterchallenged.Themortality,morbidityandprotectiverateineachgroupwascalculatedaccord-ingtotheformula:mortality(%)=thenumberofdeadchickenuptoday15af

28、terchallenge(D15)/thenumberofsample×100%,morbidity(%)=thenumberofchickensdeadandshowingclinicalsymptomsonD15/thenumberofsample×100%,protectiverate(%)=thenumberofchickenswithoutclinicalsymptomsonD15/thenumberofs

29、ample×100%.2.6.LymphocytesproliferationassayLymphocytesproliferationwasperformedaspreviouslydescribed(Zhaoetal.,2011).Briefly,bloodsampleswerecollectedfromheart,andthencarefullylayeredonthesurfaceoflymphocyteseparat

30、ionmedium.Aftercentrifugated,lymphocyteswerecol-lectedandwashedtwicewithRPMI-1640.Thenweredilutedinto2.5×106 mL?1 andaddedinto96-wellplateswith80?Lwell?1,another20?LofPHAwasadded.Theplateswereincubatedinahumidatmosp

31、hereof5%CO2 at37.5 ?C.Afteraculturefor44h,20?LofMTT(5?gmL?1)wasadded,andtheplateswerere-incubatedfor4h.Thenthesupernatantwasremovedand100?LofDMSOwereaddedandshakenfor5min.Theabsorbanceat570nm(A570 value)wasmeasuredasthei

32、ndexofperipheralTlymphocytesproliferation.2.7.SerumHIantibodyassayBloodsamplewerecollected,putintoEppendorftubesandallowedtoclotat37 ?Cfor2h.SerumwasseparatedandinactivatedcomplementsforassayingHIantibody.Themethodwasref

33、erredtothepreviousreport(Zhaoetal.,2011).Themeantiterwasexpressedasreciprocallog2 valuesofthehighestdilutionthatdisplayedHI.2.8.MeasurementofCD4+ andCD8+ TcellsTlymphocyteswerecollectedaccordingtothemethodin2.5.Then50?Lo

34、flymphocyteswereaddedtoFalcontubes.Afteradded2?Lofmonoclonalantibodies(anti-CD4+-FITCandanti-CD8+-PE),themixturewasincubatedinthedarkat4 ?Cfor30min.Aftercen-trifugation(2000×g,at4 ?Cfor5min),thecellswerewashedwithPB

35、S,andresuspendedin0.5mLofPBS.ThepercentagesofCD4+ andCD8+ Tcellsweremeasuredbyflowcytometry(Yang,Chen,Sun,Ye,&Fang,2007).2.9.StatisticalanalysisDataareexpressedasthemean±S.D.Duncan’smultiplerangetestwasusedtodet

36、erminethedifferencesamonggroupswiththesoftwareSPSS19.0.?2-Testwasusedtoanalyzethedifferenceofthemortality,morbidityandprotectiverate.SignificantdifferenceswereconsideredasP<0.05.3.Results3.1.Non-specificimmuneresponse

37、experiment3.1.1.TheeffectofOPLonphagocytosisofmacrophagesTheeffectofOPLonphagocytosisofneutralredisillustratedinFig.1.At125–7.813?gmL?1,theA490 valuesinOPLandOPgroupsweresignificantlyhigherthanthoseinBLandCCgroup(P<0.

38、05).TheA490 valuesinOPLat125,31.25and7.813?gmL?1 groupsweresignificantlyhigherthanthoseinallOPgroups(P<0.05).3.1.2.TheeffectofOPLonIL-2secretionTheeffectofOPLonthereleaseofIL-2frommecrophagesisshowninFig.2.TheIL-2conc

39、entrationsinOPLat125–15.625?gmL?1 andOPat125–31.25?gmL?1 groupsweresignificantlyhigherthanthoseinBLandCCgroups(P<0.05).TheIL-6concentrationsinOPLweresignificantlyhigherthanthoseinOPat125–31.25?gmL?1 groups(P<0.05).