肺炎克雷伯杆菌特异性卵黄抗体的制备与活性的的研究

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1、结论：实验中采用的分离纯化方法可获得高纯度的IgY。所制备的特异性IgY具有高效价和明显的体外抑菌活性。特异性IgY对小鼠肺炎克雷伯杆菌肺炎有保护作用。关键词：特异性卵黄抗体肺炎克雷伯杆菌肺炎PreparationandEfficacyEvaluationofEggYolkImmunoglobulin(IgⅥagainstKlebsiellaPneumoniaeMasterdegreecandidate：Fan—LikongSupervisor：ProfessorYu—HongZhenMajor：PharmacologyAbstractObjective

2、：Topreparespecificchickeneggyolkimmunoglobulin(ig矽againstKlebsiellapneumoniae(K．pneumonia)，evaluateitsactivityinvitroandinvivo．Methods：SpecificIgYwasproducedbyimmunizinghens、析tllformaldehydekilledKpneumoniaandpurifiedbywaterdilution，saltprecipitationsandultrafiltration．Thepuritya

3、ndtiterofIgYweredetectedbySDS-PAGEandELISA，respectively．GrowthinhibitionofspecificIgYatdifferentconcentrationstoKpneumoniawastestedinliquidculturemedium．PneumoniamodelWasestablishedbynasalinstillationofKpneumonia(2x1011cfu／m1)atadoseof10091．Micewererandomlydividedintosixgroupsand

4、protecteddifferently：blankcontrolgroup(nochallenge，)，negativecontrolgroup(normalsaline)，cefoperazonegroup(1mg／m1)，dexamethasonegroup(O．5mg／m1)，highdosespecificIgYgroup(40mg／m1)，lowdosespecificIgYgroupOomg／m1)．Micewereintraperitoneallyinjectedfrom1-daybeforechallengeto4-dayafterch

5、allengeatanintervalof24hwiththefollowingdoses(per109weight)：dexamethasonegroup，100山beforechallengeand150山afterchallenge；othergroups，150山beforechallengeand250肛lafterchallenge．Clinicalmanifestationsofmicewereobserved，thechangesoftheirweight，temperature，whitebloodcell(wBc)，deathrate

6、s，bacterialinlungtissue，appearanceoflungtissueandpulmonarypathologywereestimated．Results：ThepurityofIgYobtainedbywaterdilution，saltprecipitationsandultrafiltmtionwassimilar、析t11thatofstandardIgY．ELISAindicatedthatthetiterofspecificIgYreachedtopickvalue1600Oil40-45dayafterthefirst

7、immunization．SpecificIgYatconcentrationsof1—20mg／mlirflfibitedthegrowthofK．pneumoniaina3dosedependentmanner．AfterinfectedwithKpneumonia,thebodytemperature，weightandWBCinbloodofmicedecreasedobviouslyinallgroupsexcepttheblankcontrolgroup．ButthechangesofIgYgroupandpositivecontrol(de

8、xamethasoneandcefoperazone)groupswereles