Mucosal T Cells: Is Tolerance Floating on Lipid Rafts

粘膜 T 细胞：耐受性是否漂浮在脂筏上

基本信息

批准号：
7163549
负责人：
Alan David Levine
金额：
$ 25.39万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-01-15 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7163549
关键词：
Actins Adaptor Signaling Protein Affect Antigens Attention Authorization documentation Biochemical Blood CD2-associated protein CD3 Antigens Cell membrane Cholesterol Chronic Cities Complex Confocal Microscopy Coupling Crohn&apos s disease Cytoskeleton Disclosure Doctor of Medicine Doctor of Philosophy Eating Electrons Employee Strikes Equilibrium Event Exclusion Face Flow Cytometry Funding Gastrointestinal tract structure Generations Heterogeneity Homeostasis Immune Immune Tolerance Immune response Immune system Immunity Immunologic Factors Immunologic Techniques Inflammation Inflammatory Bowel Diseases Instruction Intermediate Filaments Intestines Investigation Ions Kinetics Lamina Propria Localized Mediating Membrane Membrane Microdomains Microbial Genetics Modeling Molecular Mucosal Immunity Mucous Membrane Names Numbers Ohio Outcome Pathogenesis Pathway interactions Peripheral Phosphoric Monoester Hydrolases Physiological Principal Investigator Property Protein Tyrosine Kinase Protein phosphatase Proteins Receptor Activation Receptor Signaling Regulation Relative (related person)Research Personnel Research Project Grants Role Second Messenger Systems Signal Transduction Signal Transduction Pathway Signaling Protein Sphingolipids Sting Injury Structure T-Cell Activation T-Cell Receptor T-Lymphocyte Techniques Testing Tissues Tweens Tyrosine Phosphorylation Ulcerative Colitis Universities Washington concept in vitro Model medical schools molecular imaging multidisciplinary novel peripheral blood programs receptor response second messenger

项目摘要

Investigations on the regulation of normal intestinal immunity have described a fundamental property of the intestinal immune system that distinguishes it from other peripheral and tissue responses. "Physiological inflammation" is a state of immune[ regulation in the intestine characterized by hypo-responsive T lymphocytes. This unique, but molecularly undefined _roperty indicates that active tolerance is an essential component of mucosal immunity. We propose that regulated taucosal T cell hypo-responsiveness mediates intestinal immune tolerance, and that unregulated T cell activation i fitiates inflammatory bowel disease (IBD). In striking contrast to peripherial blood T cells (PBT), which are activated vi a the CD3 pathway, intestinal lamina propria T cell (LPT) responses are dominated by an alternate pathway that uses the CD2 receptor. We have developed an in vitro model in which LPT can toggle between a hypo-responsive (tolerant) s:ate and a responsive (protective) state of activation. Using this model we demonstrated that membrane proximal signaling within 2 minutes of activation through the CD3 receptor complex is markedly reduced in tolerant, but not pro ective LPT. We have also described that cholesterol-rich membrane microdomains, called lipid rafts, are not only structtLral components of the plasma membrane, but also localize proteins for the initiation of intra-cellular signal transductio: in PBT. With our expertise in characterizing LPT, isolating lipid rafts, and delineating signal transduction pathways iJ T cells, we are immediately poised to study mucosal T cell tolerance in the normal mucosa and aberrant T cell activat on in the IBD mucosa at a molecular level. Our ability to identify and characterize heterogeneity among lipid raft ct,nstituents in LPT will be combined with molecular, biochemical, and immunological techniques to test the following _:entral hypothesis: Lipid raft heterogeneity on intestinal T cells modulates their signaling, activation, and functi)n, thereby contributing to both normal mucosal immune tolerance and chronic inflammation. Our aims are a, follows: (1) Evaluate the structural mechanisms by which lipid raft heterogeneity modulates positive and negative rq_gulation of the TCR signaling complex in tolerant LPT. (2) Investigate the topological coupling of the cytoskeleton to lipid rafts via interactions through CD2 in LPT. (3) Explore the biochemical regulation of the second messenger Ras-MAPK pathway in lipid rafts from LPT. (4) Examine changes in lipid raft modulation of T cell activation from IBD mucosa. We believe this unique approach will generate important new information on intestinal immune tolerance and IBD, by examining the regulation of membrane proximal events in LPT signaling and how these events chaqge in the IBD T cell. PERFORMANCESITE(S) (organization,city, state) I Case We_stern Reserve University School of Medicine Cleveland, Ohio KEYPERSONNEL.Seeinstructions. Usecontinuationpagesas neededto providethe requiredinformationin theformat shownbelow. Startwith PrincipalInvestigator.Listall otherkey personnelinalphabeticalorder,lastnamefirst. Name Organization RoleonProject Levine, Alan D., Ph.D. Case Western Reserve University P.I. Brady-Kalnay, Susan, Ph.D Case Western Reserve University Consultant Brown, Deborah, Ph.D. State University NY at Stony Brook Consultant Eppell, Steven, Ph.D. Case Western Reserve University Co-Investigator Fiocchi, Claudio, M.D. Case Western Reserve University Co-Investigator Jacobberger, James, Ph.D. Case Western Reserve University Co-Investigator Nieminen, Anna-Liisa, Ph.D. Case Western Reserve University Co-Investigator Pilar, Guillermo Ph.D. Case Western Reserve University Consultant Shaw, Andrey, M.D. Washington University, St. Louis, MO Consultant ! Disclosure Permission Statement. Applicable to SBIR/STTR Only. See instructions. [] Yes [] No / PHS398(Rev.05/01) Page 2 FormPage2 Use1/2-inchMARGINS.Numberpagesconsecutivelyat the bottomthroughoutheapplication.Dono_uttsesuffixessuchas 3a, 3b. PrincipalInvestigator/ProgramDirector(Last,first,middle): Levine, Alan D. Thenameof theprincipalinvestigator/programdirectormust beprovidedat thetop of eachprintedpageandeach continuationpage. Typedensityandsizemustconformto limits andspecificationsprovidedinthe PNS398 Instructions. RESEARCH GRANT TABLE OF CONTENTS PageNumbers Face Page ......................................................................................................................................................... 1 Description,

对正常肠道免疫调节的调查已经描述了肠道的基本特性免疫系统将其与其他外围和组织反应区分开。 “生理炎症”是一个状态免疫[以低反应性T淋巴细胞为特征的肠道调节。这个独特但分子未定义的_Roperty表明主动耐受性是粘膜免疫的重要组成部分。我们提出了这一点调节的taucosal T细胞低反应性介导肠道免疫耐受性，并且不调节的T细胞激活I适合炎症性肠病（IBD）。与周围血液T细胞（PBT）形成鲜明对比的是激活的VI A CD3途径，肠道层植物T细胞（LPT）响应由替代途径主导使用CD2受体。我们已经开发了一个体外模型，其中LPT可以在低反应性之间切换（耐受）S：Ate和一种响应性（保护性）激活状态。使用此模型，我们证明了膜通过CD3受体复合物激活后2分钟，近端信号传导显着降低，耐受性，但不是支持LPT。我们还描述了富含胆固醇的膜微区域，称为脂质筏，不是仅质膜的结构成分，但也将蛋白质定位用于启动细胞内信号 thranductio：在PBT中。我们在表征LPT的专业知识，隔离脂质筏和描述信号转导方面的专业知识途径IJ T细胞，我们立即准备研究正常粘膜和异常T的粘膜T细胞耐受性分子水平的IBD粘膜中的细胞活动。我们识别和表征异质性的能力 LPT中的NSTITENTS将与分子，生化和免疫学技术结合使用脂质筏CT，以测试以下是_：肠道假设：肠道T细胞上的脂质筏异质性调节其信号传导，激活， n功能，从而有助于正常的粘膜免疫耐受性和慢性炎症。我们的目的是a，遵循：（1）评估脂质筏异质性调节阳性和阳性的结构机制耐受性LPT中TCR信号复合物的负rq_gulation。（2）研究细胞骨架通过LPT中的CD2通过相互作用到脂质筏。（3）探索第二个 LPT的脂质筏中的Messenger Ras-Mapk途径。（4）检查T细胞脂质筏调节的变化 IBD粘膜的激活。我们认为，这种独特的方法将产生有关肠道的重要新信息免疫耐受性和IBD，通过检查LPT信号中膜近端事件的调节以及如何调节 IBD T细胞中的事件。表演场（S）（组织，城市，州）我案例We_stern储备大学医学院俄亥俄州克利夫兰 Kepersonnel.SeeeInstructions。 usecontinuationPagesas需要在format示出的below中提供必要的信息。 startwith princtionalInvestigator.listall其他关键人员人事按钮订单，lastnamefirst。名称组织角色注射器 Levine，Alan D.，博士案例西部储备大学P.I. Brady-Kalnay，Susan，博士案例西部储备大学顾问布朗，黛博拉博士Stony Brook顾问纽约州立大学 Eppell，Steven，Ph.D。案例西部储备大学共同投资者 Fiocchi，Claudio，医学博士Case Western Reserve University共同投资者 Jacobberger，James，Ph.D。案例西部储备大学共同投资者 Nieminen，Anna-Liisa博士案例西部储备大学共同投资者 Pilar，Guillermo博士案例西部储备大学顾问肖，安德烈，医学博士华盛顿大学，圣路易斯，密苏里州顾问呢披露许可声明。仅适用于SBIR/STTR。请参阅说明。 []是[]否 / PHS398（Rev.05/01）第2页FormPage2 use1/2-英寸玛金。校长申请员/程序导演（最后，第一，中间）：莱文，艾伦·D。 principalinvestigator/programDirectormust beprovidedat thetop thetop thetop theTop。 typedsentyandsizemustConformto限制和规范化的PNS398说明。研究赠款目录武器脸部页面.......................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................... 描述，