Mucosal T Cells: Is Tolerance Floating on Lipid Rafts

对正常肠道免疫调节的研究已经描述了肠道免疫的基本性质， 免疫系统将其与其他外周和组织反应区分开来。“生理性炎症”是一种 肠道内的免疫[调节]，以低反应性T淋巴细胞为特征。这种独特的，但分子 undefined _roperty表明主动耐受是粘膜免疫的基本组成部分。我们建议 受调节的Tauclitaxin T细胞低反应性介导肠道免疫耐受，而未受调节的T细胞低反应性介导肠道免疫耐受， 激活导致炎症性肠病（IBD）。与外周血T细胞（PBT）形成鲜明对比的是， 肠固有层T细胞（LPT）通过CD 3途径激活，其应答由另一途径主导 它使用CD 2受体。我们已经开发了一种体外模型，其中LPT可以在低反应和低反应之间切换。 （耐受性）和响应性（保护性）激活状态。利用这个模型，我们证明了膜 在耐受性， 但不是预期的LPT。我们还描述了富含胆固醇的膜微区，称为脂筏， 不仅是质膜的结构成分，而且还定位用于启动细胞内信号的蛋白质 transductio：在PBT中。凭借我们在表征LPT，分离脂筏和描绘信号转导方面的专业知识， 通过研究T细胞的免疫途径，我们立即准备研究正常粘膜和异常T细胞中的粘膜T细胞耐受性。 细胞在分子水平上活化。我们识别和描述 脂筏CT，LPT中的成分将结合分子，生化和免疫学技术来检测LPT中的脂筏CT， 以下是中心假说：肠T细胞上的脂筏异质性调节它们的信号传导，活化， 和Functin，从而有助于正常粘膜免疫耐受和慢性炎症。我们 目的是：（1）评价脂筏异质性调节正性和负性的结构机制， 在耐受性LPT中TCR信号复合物的负调节。(2)研究的拓扑耦合 细胞骨架通过LPT中的CD 2相互作用形成脂筏。(3)探索第二种生化调控 LPT脂筏中的信使Ras-MAPK通路。(4)检测T细胞脂筏调节的变化 从IBD粘膜激活。我们相信这种独特的方法将产生重要的新信息， 免疫耐受和IBD，通过检查LPT信号传导中的膜近端事件的调节以及这些事件如何影响免疫耐受和IBD。 IBD T细胞中的事件。 地点（组织、城市、州） 我 案例我们_斯特恩储备大学医学院 克利夫兰，俄亥俄州 KEYPERSONNEL.Seeinstructions.需要使用第二个续页以下面所示的格式提供所需信息。 从主要研究者开始，按顺序列出其他关键人员，姓氏在前。 名称组织机构RoleonProject 莱文，艾伦D.，博士凯斯西储大学P.I. Brady-Kalnay，Susan，博士凯斯西储大学顾问 布朗，黛博拉，博士州立大学纽约斯托尼布鲁克分校顾问 Eppell，Steven，博士凯斯西储大学共同研究员 Fiocchi，Claudio，医学博士凯斯西储大学共同研究员 Jacobberger，James，博士凯斯西储大学共同研究员 Nieminen，Anna-Liisa，博士凯斯西储大学共同研究员 皮拉尔，吉列尔莫博士凯斯西储大学顾问 Shaw，Andrey，医学博士密苏里州圣路易斯华盛顿大学 ! 披露许可声明。仅适用于SBIR/STTR。参见说明。[]是[]否 / PHS 398（修订版05/01）第2页表格第2页 1/2英寸的边距。编号页在整个应用程序的底部。请不要修改3a，3b。 主要研究者/项目负责人（最后，第一，中间）：Levine，Alan D. 主要研究者/项目负责人的姓名必须在打印的每页和每个续页的顶部提供。 类型密度和尺寸必须符合PNS 398说明中提供的限值和规格。 研究资助 目录 页码 首页......................................................................................................................................................... 1 说明，

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,