会员注册 | 登录 | 微信快捷登录 支付宝快捷登录 QQ登录 微博登录 | 帮助中心 人人文库renrendoc.com美如初恋!
站内搜索 百度文库

热门搜索: 直缝焊接机 矿井提升机 循环球式转向器图纸 机器人手爪发展史 管道机器人dwg 动平衡试验台设计

   首页 人人文库网 > 资源分类 > PDF文档下载

共刺激分子B7H3在支气管哮喘中的作用研究

  • 资源星级:
  • 资源大小:2.73MB   全文页数:58页
  • 资源格式: PDF        下载权限:注册会员/VIP会员
您还没有登陆,请先登录。登陆后即可下载此文档。
  合作网站登录: 微信快捷登录 支付宝快捷登录   QQ登录   微博登录
友情提示
2:本站资源不支持迅雷下载,请使用浏览器直接下载(不支持QQ浏览器)
3:本站资源下载后的文档和图纸-无水印,预览文档经过压缩,下载后原文更清晰   

共刺激分子B7H3在支气管哮喘中的作用研究

硕士学位论文论文题目共刺激分子B7H3在支气管哮喘中的作用研究研究生姓名陈赢指导教师姓名季伟(教授)专业名称儿科学研究方向呼吸论文提交日期2013年4月共刺激分子B7H3在支气管哮喘中的作用研究中文摘要I共刺激分子B7H3在支气管哮喘中的作用研究中文摘要第一部分可溶性B7H3在支气管哮喘患儿血浆中的表达及临床意义目的研究支气管哮喘患儿外周血中可溶性B7H3(sB7H3)的表达水平,并分析其与细胞因子水平的相关性,探讨sB7H3的异常表达在支气管哮喘急性发作中的临床意义。方法选取2012年1月~2012年12月在苏州大学附属儿童医院呼吸科住院的支气管哮喘患儿21例,并选取外科择期手术(腹股沟斜疝、血管瘤、腺样体肥大)而住院的患儿16例作为对照组儿童,分为支气管哮喘急性发作期组(A组)、恢复期组(B组)及对照组(C组)。同时采集支气管哮喘患儿急性发作期及恢复期的外周血,以及外科择期手术患儿的外周血,采用酶联免疫吸附试验(ELISA)检测血浆中sB7H3、IFNγ、IL4及IL10的表达水平,及分析支气管哮喘患儿急性发作期血浆中sB7H3的水平与细胞因子IFNγ、IL4、IL10的相关性。结果哮喘急性发作期组血浆中sB7H3的水平明显高于恢复期组及对照组(P0.05),而恢复期组虽高于对照组,但无统计学差异哮喘急性发作期组血浆中IFNγ、IL10水平均低于恢复期组及对照组(P0.05),恢复期与对照组相比无统计学差异血浆中IL4的水平在哮喘急性发作期组与恢复期组均高于对照组(P0.05),且哮喘急性发作期组的表达水平高于恢复期组(P0.05)。支气管哮喘患儿急性发作期血浆中sB7H3的表达水平与IL4的水平具有正相关性,与IFNγ、IL10的水平无相关性。结论sB7H3在支气管哮喘急性发作期异常高表达sB7H3可能通过增强Th2细胞因子的分泌而在支气管哮喘的急性发作期起作用。关键词sB7H3支气管哮喘Th2细胞细胞因子中文摘要共刺激分子B7H3在支气管哮喘中的作用研究II第二部分抗B7H3单克隆抗体对哮喘小鼠模型的作用研究目的探讨抗B7H3单克隆抗体对哮喘小鼠模型诱导期及效应期的治疗作用。方法68周龄雌性BALB/c小鼠(1820g),随机均分为5组,每组10只,分别为哮喘组(A组),生理盐水对照组(B组),IgG组(C组),抗B7H3抗体诱导期组(D组),抗B7H3抗体效应期组(E组)。A、C、D、E组均给予OVA致敏及激发(0、10天给予OVA腹腔注射致敏,第20天给予OVA雾化吸入,每天半小时连续7天),C组、D组在第2,7,11,14,18天分别给予腹腔注射IgG、抗B7H3抗体,E组在第2,7,11,14,18天给予腹腔注射IgG,第21,24,26天腹腔注射抗B7H3抗体(OVA雾化1小时前注射)。B组给予同剂量生理盐水注射、激发。各组小鼠于末次激发2小时后麻醉处死,采集标本。小鼠进行肺泡灌洗,肺泡灌洗液(BALF)进行细胞总数计数及分类计数并采用ELISA测定小鼠BALF中的IFNγ、IL4、IL17的水平小鼠肺组织切片行HE染色、PAS染色及免疫组化法(SABC法),观察肺组织炎性细胞及嗜酸性细胞浸润情况、气道粘液分泌情况及B7H3表达情况。结果哮喘组肺泡灌洗液(BALF)中的细胞总数及嗜酸性细胞数多于生理盐水对照组及抗B7H3抗体诱导期组,差异具有统计学意义(P0.05),哮喘组、IgG组、抗B7H3抗体效应期组BALF中细胞总数及嗜酸性细胞数比较差异无统计学意义,抗B7H3抗体诱导期组BALF中的细胞总数及嗜酸性细胞数高于生理盐水对照组,差异具有统计学意义(P0.05)。哮喘组、IgG组、抗B7H3抗体诱导期组及抗B7H3抗体效应期组BALF中的IFNγ的水平均低于生理盐水对照组(P0.05),哮喘组、IgG组、抗B7H3抗体效应期组IFNγ的水平无统计学差异,抗B7H3抗体诱导期组IFNγ的水平高于IgG组(P0.05)。哮喘组、IgG组、抗B7H3抗体诱导期组及抗B7H3抗体效应期组BALF中的IL4、IL17的水平均高于生理盐水对照组(P0.05),哮喘组、IgG组、抗B7H3抗体效应期组IL4、IL17水平无统计学差异,抗B7H3抗体诱导期组IL4、IL17水平低于IgG组(P0.05)。肺组织病理染色显示哮喘组、IgG组、抗B7H3抗体效应期组炎性细胞、嗜酸性细胞浸润及粘液分泌较抗B7H3抗体诱导期组明显,生理盐水对照组无明显炎性细胞、嗜酸性细胞浸共刺激分子B7H3在支气管哮喘中的作用研究中文摘要III润、粘液分泌。免疫组化显示哮喘组、IgG组B7H3高表达,生理盐水对照组B7H3不表达,抗B7H3抗体诱导期组、抗B7H3抗体效应期组B7H3部分表达。结论哮喘的诱导期给予抗B7H3单克隆抗体治疗能通过抑制Th2型细胞因子(IL4)和Th17型细胞因子(IL17)的分泌,同时促进Th1型细胞因子(IFNγ)的分泌,从而减轻OVA致敏哮喘小鼠的气道病理改变。关键词抗B7H3单克隆抗体哮喘小鼠模型细胞因子作者陈赢指导老师季伟AbstractRoleofcostimulatorymoleculeB7H3inbronchialasthmaIVRoleofcostimulatorymoleculeB7H3inbronchialasthmaAbstractExpressionandclinicalsignificanceofthesolubleB7H3inplasmaofchildrenwithbronchialasthmaObjectiveTodiscussthelevelofsolubleB7H3intheperipheralbloodofchildrenwithbronchialasthma,andanalyzeitscorrelationwiththeexpressionofcytokines,toexploretheclinicalsignificanceofabnormalexpressionofsolubleB7H3inacutephaseofbronchialasthma.Methods21casesofchildrenwithbronchialasthmawereenrolledatdepartmentofrespirationofChildrenshospitalaffiliatedinSoochowuniversityfromJanuary2012toDecember2012,and16casesofchildrenIndirectinguinalhernia,hemangioma,adenoidhypertrophyhospitalizedinsurgeryascontrolgroupchildren,dividedintoacutephaseofasthmagroupgroupA,recoveryphasegroupgroupBandcontrolgroupgroupC.Atthesametime,peripheralbloodwerecollectedinchildrenwithbronchialasthmainacutephaseandrecoveryphaseaswellaschildreninsurgery,EnzymelinkedimmunosorbentassaywereusedtodetectthelevelofsolubleB7H3,IFNgamma,interleukin4andinterleukin10inplasma,andanalyzethecorrelationbetweenthelevelofsolubleB7H3andcytokinesIFNgamma,interleukin4,interleukin10inplasmainacutephaseofasthma.ResultsThelevelofsolubleB7H3inplasmainacutephaseofasthmagroupweresignificantlyhigherthantherecoveryphasegroupandcontrolgroupP0.05,whilerecoveryphasegroupwashigherthancontrolgroup,butnosignificantdifference,thelevelofIFNgammaandinterleukin10inacutephaseofasthmagroupwerelowerthanthatofrecoveryphasegroupandcontrolgroupP0.05,recoveryphasegroupandcontrolgrouphadnosignificantdifference.Thelevelofinterleukin4inplasmabothinacutephaseofasthmagroupandrecoveryphasegroupwerehigherthanthatofcontrolgroupP0.05,andthelevelinacutephaseofasthmagroupwashigherthanthatofrecoveryphasegroupRoleofcostimulatorymoleculeB7H3inbronchialasthmaAbstractVP0.05.ThelevelofsolubleB7H3andthelevelofinterleukin4inplasmahaspositivecorrelationinchildreninacutephaseofbronchialasthma,andhasnocorrelationwithIFNgamma,interleukin10.ConculsionThelevelofsolubleB7H3wasabnormalhighexpressioninacutephaseofbronchialasthma,solubleB7H3mayenhancethesecretionofTh2typecytokinesandhaseffectinacutephaseofbronchialasthma.KeywordssolubleB7H3bronchialasthmaTh2cellcytokineRoleofantiB7H3monoclonalantibodyinmousemodelofasthmaObjectiveToexplorethetherapeuticfunctionofantiB7H3monoclonalantibodyinmousemodelofasthmaininductionphaseandeffectorphase.MethodsFemaleBALB/cmiceweight1820gand68wkoldweredividedintofivegroups,everygrouphas10mice,asthmaticgroupgroupA,normalsalinecontrolgroupgroupB,IgGgroupgroupC,antiB7H3antibodyinductiongroupgroupD,antiB7H3antibodyeffectorgroupgroupE.MiceingroupA,groupC,groupDandgroupEweregivenOVAsensitizationandexcitationmiceweresensitizedtoOVAbyintraperitonealinjectionondays0and10,thentheywereexposedtoOVAaerosolchallengesforthirtyminutesonconsecutivesevendaysonday20,groupCandgroupDweregivenintraperitonealinjectionofIgGandantiB7H3antibodyondays2,7,11,14,18,groupEweregivenintraperitonealinjectionofIgGondays2,7,11,14,18,intraperitonealinjectionofantiB7H3antibodyondays21,24,26intraperitonealinjectionbeforeonehourofOVAchallenges.MiceingroupBweregivensalineasabovemethods.Aftertwohouroflastchallengeallmicewereanesthetizedtodeathtocollectsamples,allmicewerealveolaredlavage,andbronchoalveolarlavagefluidBALFwerecollectedtocountcellsnumberandclassifycells,measurethelevelofIFNgamma,interleukin4,interleukin17inBALFbyEnzymelinkedimmunosorbentassay,mouselungtissuesectionsstainedwithHE,PASandimmunohistochemicalmethodSABCmethod,toobservetheinfiltrationofinflammatorycellsandeosinophiliccells,mucussecretionandtheexpressionofB7H3inlungtissue.ResultsThetotalcellsandeosinophiliccellsinbronchoalveolarlavagefluidinasthmaticgroupweremorethanthenormalsalinecontrolgroupandantiB7H3antibodyAbstractRoleofcostimulatorymoleculeB7H3inbronchialasthmaVIinductiongroup,thedifferencewerestatisticallysignificantP0.05.Thetotalcellsandeosinophiliccellsinbronchoalveolarlavagefluidinasthmaticgroup,IgGgroupandantiB7H3antibodyeffectorgroupwerenostatisticallysignificant.ThetotalcellsandeosinophiliccellsinantiB7H3antibodyinductiontreatmentgroupweremorethanthenormalsalinecontrolgroupP0.05.ThelevelofIFNgammainBALFinasthmaticgroup,IgGgroup,antiB7H3antibodyinductiongroupandantiB7H3antibodyeffectorgroupwerelowerthanthenormalsalinecontrolgroupP0.05,thelevelofIFNgammainasthmaticgroup,IgGgroupandantiB7H3antibodyeffectorgroupwerenostatisticallysignificant,thelevelofIFNgammainantiB7H3antibodyinductiongroupwerehigherthanIgGgroupP0.05.Thelevelofinterleukin4andinterleukin17inasthmaticgroup,IgGgroup,antiB7H3antibodyinductiongroupandantiB7H3antibodyeffectorgroupwerehigherthanthenormalsalinecontrolP0.05,thelevelofinterleukin4,interleukin17inasthmaticgroup,IgGgroupandantiB7H3antibodyeffectorgroupwerenostatisticallysignificant,thelevelofinterleukin4,interleukin17inantiB7H3antibodyinductiongroupwerelowerthanIgGgroupP0.05.Lungtissuepathologicalstainedshowedmoreinfiltrationofinflammatorycells,eosinophiliccellandmucussecretioninasthmaticgroup,IgGgroup,antiB7H3antibodyeffectorgroupthanthatofantiB7H3antibodyinductiongroup,normalsalinecontrolgroupnosignificantinfiltrationofinflammatorycells,eosinophiliccellandmucussecretion.ImmunohistochemistryshowedhighexpressionofB7H3inasthmaticgroupandIgGgroup,normalsalinecontrolgroupdidnotexpressB7H3,andB7H3partlyexpressioninantiB7H3antibodyinductiongroupandantiB7H3antibodyeffectorgroup.ConculsionTreatmentwithantiB7H3monoclonalantibodyininductionphaseofasthmacaninhibitsecretionofTh2typecytokineinterleukin4andTh17typecytokinesinterleukin17,andpromotesecretionofTh1typecytokineIFNgamma,therebyreducingtheairwaypathologicalchangeofmousemodelofasthmainducedbyOVA.KeywordsAntiB7H3monoclonalantibodymousemodelofasthmacytokineWrittenbyChenYingSupervisedbyJiWei目录前言............................................................................................................................1第一部分可溶性B7H3(sB7H3)在支气管哮喘患儿血浆中的表达及临床意义...6材料与方法................................................................................................................7结果.................................................................................................................10讨论.................................................................................................................12参考文献.................................................................................................................15第二部分抗B7H3单克隆抗体对哮喘小鼠模型的作用研究...................................18材料与方法..............................................................................................................18结果.................................................................................................................23讨论.................................................................................................................29参考文献.................................................................................................................33结论........................................................................................................................35综述共刺激分子B7H3的生物学作用及与支气管哮喘的研究进展...............36缩略词表........................................................................................................................45攻读硕士期间公开发表的论文......................................................................................47致谢........................................................................................................................48共刺激分子B7H3在支气管哮喘中的作用研究前言1前言支气管哮喘(bronchialasthma),简称哮喘,是儿童最常见的气道慢性炎症性疾病,发病率在发达国家可达33,且有持续上升的趋势,粗略估计中国有3000万哮喘患者,中国014岁儿童哮喘患病率1990年到2000年从0.78上升到2.06,增加了1.6倍,有逐年上升的趋势1,支气管哮喘已经成为全球关注的公共卫生问题,从哮喘发病机制水平寻找干预措施是国内外医学界所关注的课题。支气管哮喘是以慢性气道炎症及广泛的嗜酸性细胞浸润为特点,主要表现为支气管痉挛、管壁炎症性肿胀、细支气管粘液分泌增加,各种特应性及非特应性刺激引起气道高反应及气道重塑。哮喘的发病机制极为复杂,与免疫、神经、内分泌、遗传、环境等因素密切相关,而免疫学因素被认为是哮喘最重要的作用机制,经典的免疫学说认为哮喘的发病与Th1/Th2的平衡紊乱密切相关,致敏原作用于支气管上皮细胞,促使上皮细胞分泌胸腺基质淋巴细胞生成素(thymicstromallymphopoietin,TSLP)并作用于树突状细胞(dendriticcells,DCs)表面的TSLP受体,使得DCs协同刺激分子表达增加,从而使Th0细胞向Th2型细胞分化,表现为Th2型细胞数量增多或功能亢进,从而促使IgE和IL4、IL5、IL13等细胞因子产生增多,这些细胞因子促使嗜酸性细胞、肥大细胞、T淋巴细胞等炎症细胞募集和活化,产生慢性气道炎症,同时调节气道粘液的分泌,引起气道阻塞24。调节性T细胞(Treg)也参与了哮喘的发生发展,在诱导和维持对致敏原的免疫耐受中发挥重要作用。而IL10可以由Treg分泌,且有研究表明IL10可以抑制致敏原诱导的Th2型免疫反应及IL4的分泌而阻断哮喘的发生发展26。Th17为近年新发现的Th细胞亚群,IL17为其分泌的特征性细胞因子,IL17的水平与哮喘气道高反应及哮喘严重程度呈正相关56。Th17促进中性粒细胞募集和活化,并能刺激IL6、TNFα的细胞因子的分泌,参与固有免疫应答及适应性免疫应答7。Th细胞的分化是建立在T细胞活化的基础上的,T细胞的活化及免疫应答的启动需要双信号协同作用TCR识别MHC抗原复合物,获得第一信号后,还必需获得协同信号分子提供的第二信号,即抗原呈递细胞(APC)给予的协同刺激信号,其中

注意事项

本文(共刺激分子B7H3在支气管哮喘中的作用研究)为本站会员(vyyolyg827)主动上传,人人文库网仅提供信息存储空间,仅对用户上传内容的表现方式做保护处理,对上载内容本身不做任何修改或编辑。 若此文所含内容侵犯了您的版权或隐私,请立即通知人人文库网([email protected]),我们立即给予删除!

温馨提示:如果因为网速或其他原因下载失败请重新下载,重复下载不扣分。

copyright@ 2015-2017 人人文库网网站版权所有
苏ICP备12009002号-5