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CT联合ESA鼻内免疫小鼠诱导的抗弓形虫感染保护性免疫应答.docx

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CT联合ESA鼻内免疫小鼠诱导的抗弓形虫感染保护性免疫应答 Introduction ToxoplasmosisisaparasiticdiseasecausedbytheprotozoanToxoplasmagondii(T.gondii),whichaffectsawiderangeofanimals,includinghumans.Theinfectionisacquiredthroughtheingestionofcontaminatedfoodorwater,orthroughcontactwithinfectedanimals.Inhumans,toxoplasmosisisusuallyasymptomatic,butitcancauseseverediseaseinimmunocompromisedindividualsandcongenitalinfectionsinpregnantwomen,leadingtofetaldeath,developmentalabnormalities,andneurologicaldeficits.Currently,thereisnoeffectivevaccineforT.gondii,butseveralexperimentalapproachesarebeingdeveloped. Onepromisingstrategyistheuseofrecombinantproteinsasantigensinasubunitvaccine.However,thesevaccinesareoftenpoorinducersofcellularimmunity,whichisessentialforcontrollingT.gondiiinfection.Toenhancetheprotectiveimmuneresponses,variousadjuvantshavebeenevaluated,includingcytokines,Toll-likereceptoragonists,andimmunemodulators. Recently,thecombineduseofcholeratoxin(CT)andEscherichiacoliheat-labileenterotoxin(LT)asmucosaladjuvantshasbeenshowntoinducepotentsystemicandmucosalimmuneresponsesinpreclinicalstudies.Inthisstudy,wetestedthehypothesisthatco-administrationofCTandLTwithT.gondiiantigenicproteinsusinganasalroutewouldinduceaprotectiveimmuneresponseinmice. MaterialsandMethods Mice Six-week-oldBALB/cfemalemice(n=24)wereobtainedfromacommercialbreeder(ShanghaiLaboratoryAnimalCo.,Ltd).Theanimalsweremaintainedunderspecific-pathogen-freeconditionsattheanimalfacilityoftheShanghaiPublicHealthClinicalCenter.TheexperimentalprotocolswereapprovedbytheAnimalEthicsCommitteeofthecenter. Antigens TherecombinantT.gondiiantigensusedinthisstudywereGRA1andSAG1,whicharewell-knownimmunodominantsurfaceantigens.TheantigenswereproducedinEscherichiacoliandpurifiedusingNi-NTAchromatographyandgelfiltration.ProteinconcentrationwasdeterminedusingtheBradfordmethod,andpuritywasconfirmedbySDS-PAGEandWesternblotting. Adjuvants Choleratoxin(CT)andEscherichiacoliheat-labileenterotoxin(LT)wereobtainedfromSigma-Aldrich.Thetoxinsweredilutedi