Uropathogenic E.coli PapG Protein: The Expression and Immune Characterization in Balb/C Mice

Journal of Applied Biological Sciences

Mohammad Reza AGHASADEGHI¹  Fatemeh ASHRAFI¹  Jalil Fallah MEHRABADI¹  Seyed Davar SIADAT¹ 

关键词: Uropathogenic E.coli;    PapG;    expression;    immunization;    immune responses;   

DOI  :  

学科分类：生物科学（综合）

来源: Nobel Bilim ve Arasirma Merkezi/Nobel Science and Research Cente

【 摘 要 】

Urinarytractinfection(UTI)isoneofthemostcomplicationsinhumancausedbyuropathogenicEscherichiacoli(UPEC)andthereisnoeffectivevaccineagainstUTI.InthisstudythePapGproteinwasexpressedandmonitoredforimmunoassay.ThepEXA/PapGvectorwasdigestedwithNdeI/EcoRIenzymes,andthegenesegmentwassub-clonedintopET21avector.AftertransformationtoE.colistrainOrigami,PapGwasexpressedunderthe1mMIPTG.ProteinexpressionwasdetectedbySDS-PAGEandwesternblotting.ProteinpurificationwascarriedoutusingNi-NTAcolumn.Experimentalmice(n=18)weredividedintothreegroups.Thegroups1to3wereimmunizedthreetimeswith2weeksintervalwiththePapGproteinwithfreund’sadjuvant,alumadjuvantedproteinandPBS,asacontrol.Twoweeksafterthelastinjection,lymphocyteproliferationassessedwithBrdumethod,secretionofIL-4andIFN-γcytokineswerequantifiedbyELISAkit.Inaddition,totalantibody,IgG1andIgG2asubtypeswereassayedwithELISA.TheresultsindicatedthatPapG,asavaccinecandidate,significantlyincreaselymphocyteproliferationandTh1cytokinepatternversuscontrolgroup.HumoralimmuneresponsesinducedwithbothIgG1andIgG2aclasses.AlthoughPapG,asavaccinecandidate,stronglyinducedcellularandhumoralimmuneresponses,morestudiesandexperimentalbacterialchallengesremainedtobeclarified.

【 授权许可】

Unknown