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Page 1: •dp»pAB…‰GHl¾¿À`´|cjpb.org/pdfup/2018042505895329.pdf · 2018-04-25 · globulindomainandmucindomain3,TIM3)B„Ø ‹Zi3ˇœß1(Programmedcelldeathprotein1, PD1)K|¥æ!,9z{Ž1。

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[犑狅狌狉狀犪犾狅犳犘犪狋犺狅犵犲狀犅犻狅犾狅犵狔.2018Mar;13(3):278-281,286.]

犃狆狉犲犾犻犿犻狀犪狉狔狊狋狌犱狔狅犳狋犺犲犿犲犮犺犪狀犻狊犿犳狅狉狉犲犱狌犮犲犱犲犳犳犻犮犪犮狔狅犳犪狀犱狆狉狅狋犲犮狋犻狅狀犫狔犪犿犪犾犪狉犻犪狏犪犮犮犻狀犲

JIANGLinping1,GUANHongli

1,TENGPingying1,MOGang

1,LIDayu1,PENGXiaohong

1,2

 (1.犇犲狆犪狉狋犿犲狀狋狅犳犘犪狉犪狊犻狋狅犾狅犵狔,犌狌犻犾犻狀犕犲犱犻犮犪犾犝狀犻狏犲狉狊犻狋狔,犌狌犻犾犻狀,犌狌犪狀犵狓犻541004,犆犺犻狀犪;2.犇犲狆犪狉狋犿犲狀狋狅犳

犘犪狋犺狅犵犲狀犅犻狅犾狅犵狔,犜犺犻狉犱犕犻犾犻狋犪狉狔犕犲犱犻犮犪犾犝狀犻狏犲狉狊犻狋狔)

【犃犫狊狋狉犪犮狋】 犗犫犼犲犮狋犻狏犲狊 Toinvestigatethemechanismforreducedefficacyofandprotectionbyamalariavaccine. 

犕犲狋犺狅犱狊 C57BL/6orBALB/cmicewereintravenouslyinoculated3timeswith10kor50kWT犘犾犪狊犿狅犱犻狌犿狔狅犲犾犻犻spo

rozoites(spz)at4weekintervals.Threehoursafterinjection,themiceweretreatedwith0.8mgofchloroquine(CQ)

for10consecutivedays.Micewerechallengedwith10kWTspzthatwereinjectedintravenously30,90,or180dafter

immunization.Thephenotypeofhepaticlymphocyteswasdeterminedwithflowcytometry. 犚犲狊狌犾狋狊 BothC57BL/6and

BALB/cmicehad100% protection30dafterimmunization.However,10% ofC57BL/6mice(1/10)haddetectable

bloodstageparasitemia90dafterimmunization,and60%ofC57BL/6mice(6/10)hadparasitemia180dafterimmuniza

tion.AlloftheBALB/cmicehad100%protection180dafterimmunization.Phenotypicanalysisoflymphocytesindica

tedthatPD1expressiononCD8+TcellsincreasedsignificantlyinC57BL/6mice90dafterimmunizationbutnotin

BALB/cmice.Moreover,thelevelofPD1expressioninmice180dafterimmunizationwassignificantlyhigherthanthat

inmice90dafterimmunization(狋=6.175,犘<0.01). 犆狅狀犮犾狌狊犻狅狀 Thereducedefficacyofandprotectionbymalaria

vaccinewascorrelatedwithahighlevelofPD1expressiononCD8+Tcells.

【犓犲狔狑狅狉犱狊】 犘犾犪狊犿狅犱犻狌犿狔狅犲犾犻犻;vaccine;CD8+Tcells;PD1

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Page 2: •dp»pAB…‰GHl¾¿À`´|cjpb.org/pdfup/2018042505895329.pdf · 2018-04-25 · globulindomainandmucindomain3,TIM3)B„Ø ‹Zi3ˇœß1(Programmedcelldeathprotein1, PD1)K|¥æ!,9z{Ž1。

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[2] ParobekCM,ParrJB,BrazeauNF,etal.Partnerdrugresist

anceandpopulationsubstructuringofartemisininresistant犘犾犪狊

犿狅犱犻狌犿犳犪犾犮犻狆犪狉狌犿inCambodia[J].GenomeBiolEvol,2017,9

(6):1673-86.

[3] BenelliG,BeierJC.Currentvectorcontrolchallengesinthefight

againstmalaria[J].ActaTrop,2017,174:91-6.

[4] SchmidtNW,HartyJT.Cuttingedge:attritionof犘犾犪狊犿狅犱犻狌犿

specificmemoryCD8Tcellsresultsindecreasedprotectionthatis

rescuedbyboosterimmunization[J].JImmunol,2011,186(7):

3836-40.

[5] NganouMakamdopK,vanGemertGJ,ArensT,etal.Long

termprotectionafterimmunizationwith犘.犫犲狉犵犺犲犻sporozoites

correlateswithsustainedIFNgammaresponsesofhepaticCD8+

memoryTcells[J].PLoSOne,2012,7(5):e36508.

[6]  ��

,öJ

,d�¡.NKZiýTLR9pØtCpGs¿�h

J9[³ñ LnKÀN

[J].n§�³¡¸&��

,2012,7

(9):675-8.

[7] Olotu,A.,FeganG,WambuaJ,etal.Fouryearefficacyof

RTS,S/AS01Eanditsinteractionwithmalariaexposure[J].N

EnglJMed,2013,368(12):1111-20. (782869

·182·

. / 0 1 2 3 4 5 6

犑狅狌狉狀犪犾狅犳犘犪狋犺狅犵犲狀犅犻狅犾狅犵狔 2018ñ3ò '13ó'3h

March2018,Vol.13,No.3

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[1] AlvarJ,VélezID,BernC,etal.Leishmaniasisworldwideand

globalestimatesofitsincidence[J].PlosOne,2012,7(5):

e35671.

[2] êÆþ

,¹ÇÈ

,± ½

,_.¦§2005-2015ñý�����

¤�R@A;ï

[J].n­�R�&��

,2017,38(4):431-4.

[3] ChuKH,LiCP,HoHY.Thefirstinternaltranscribedspacer

(ITS1)ofribosomalDNAasamolecularmarkerforphylogenetic

andpopulationanalysesincrustacea[J].MariBiotechnol,2001,

3(4):355-61.

[4] XÿS

,ùÐÉ

,ÊË./{·9J

(ITS)ýð¡ñ;?¡¸&

;�nK_NB¥Q^

[J].n§Ø¸Cj�&�

,2008,16(4):

41-7.

[5] {IÌ

,�X�

,TÍN

,_.PCR-[§L»¯°³ñ1{¥

ijKàá

[J].n§ð¡ñ&Çð¡ñ���

,2006,24(2):

92-6.

[6] NasereddinA,BensoussanHermanoE,SchonianG,etal.Mo

leculardiagnosisofOldWorldcutaneousleishmaniasisandspecies

identificationbyuseofareverselineblothybridizationassay.[J].

JClinMicrobiol,2008,46(9):2848-55.

[7] BerzunzaCruzM,CabreraN,CrippaRossiM,etal.Polymor

phismanalysisoftheinternaltranscribedspacerandsmallsubunit

ofribosomalRNAgenesof犔犲犻狊犺犿犪狀犻犪mexicana[J].Parasitol

Res,2002,88(10):918-25.

[8] �ý­

,ÎôÌ

,ºè~

,_.¦§�"6J¤9»¯°³ñ

ITS1STéêK7��;ï

[J].n§�³¡¸&��

,2010,

5(7):520-3.

[9] JamjoomMB,AshfordRW,BatesPA,etal.犔犲犻狊犺犿犪狀犻犪dono

vaniistheonlycauseofvisceralleishmaniasisinEastAfrica;pre

viousdescriptionsof犔.犻狀犳犪狀狋狌犿and"犔.犪狉犮犺犻犫犪犾犱犻"fromthis

regionareaconsequenceofconvergentevolutionintheisoenzyme

data.[J].Parasitology,2004,129(4):399-409.

[10] ºÏ

,�X�.;?}~ý»¯°³ññLÖ'u`| ¡à

ánK_N

[J].§ì+&ð¡ñ���

,2006,33(5):239-

44.

[11] éEÐ

,©ÏÑ

,¶pè

,_.$6úû"�&}~x�4»¯

°�m±¬¸V��·³KÖ'

[J].n§�³¡¸&��

2017,12(6):528-31

[12] ÎôÌ.»¯°³ñK�R�&45BYZ;\àá

[D].Ç#

%&

,Ç#

:2011.

[13] öa

,k´Ò

,± ½

,_.�ÞÀæ³´µ\ý���RJm

Ö»¯°³ñij45

[J].n§�³¡¸&��

,2015,10(3):

261-4.

[14] åÃ

,öн.¦§�"6J¤9»¯°³ñITSSTKêù

Béê;ï

[J].¡¸+&��&��

,2005,22(3):540-4.

【���n

】 20171227 【���n

】 

檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿

20180220

(}~2819

[8] RoestenbergM,TeirlinckAC,McCallMB,etal.Longtermpro

tectionagainstmalariaafterexperimentalsporozoiteinoculation:

anopenlabelfollowupstudy[J].Lancet,2011,377(9779):

1770-6.

[9] FranciscoLM,SagePT,SharpeAH.ThePD1pathwayintoler

anceandautoimmunity[J].ImmunolRev,2010,236:219-42.

[10] MartinezSalazarMB,DelgadoDomínguezJ,SilvaEstradaJ,et

al.VaccinationwithLeishmaniamexicanaLPGinducesPD1in

CD8(+)andPDL2inmacrophagestherebysuppressingtheim

muneresponse:amodeltoassessvaccineefficacy[J].Vaccine,

2014,32(11):1259-65.

[11] YoungbloodB,NotoA,PorichisF,etal.Cuttingedge:Pro

longedexposuretoHIVreinforcesapoisedepigeneticprogramfor

PD1expressioninvirusspecificCD8Tcells[J].JImmunol,

2013,191(2):540-4.

[12] NicholasK.,ZernEK,BarnettL,etal.BcellresponsestoHIV

antigenareapotentcorrelateofviremiainHIV1infectionandim

provewithPD1blockade[J].PLoSOne,2013,8(12):e84185.

[13] LuP,YoungbloodBA,AustinJW,etal.Blimp1repressesCD8

TcellexpressionofPD1usingafeedforwardtranscriptionalcir

cuitduringacuteviralinfection[J].JExpMed,2014,211(3):

515-27.

【���n

】 20171226 【���n

】 20180225

·682·

. / 0 1 2 3 4 5 6

犑狅狌狉狀犪犾狅犳犘犪狋犺狅犵犲狀犅犻狅犾狅犵狔 2018ñ3ò '13ó'3h

March2018,Vol.13,No.3