TUMOR CELL DESTRUCTION--PATH FROM BINDING TO LYSIS

肿瘤细胞破坏——从结合到裂解的路径

基本信息

批准号：
3458927
负责人：
LLOYD GRAY
金额：
$ 9.93万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-05-05 至 1994-04-30
项目状态：
已结题

项目摘要

The long term goal of this proposal is to arrive at a comprehensive understanding of the mechanism of signal transduction and lethal hit delivery in cytolytic T lymphocytes (CTL). CTL are an ultimate effector cell in the cellular arm of the immune response and are involved in host defense from neoplasia and viral illness as well as graft rejection and graft-vs-host disease. Thus, the mechanism by which these cells are activated to produce these effects is of fundamental biomedical importance. The Specific Aims of this proposal are: 1. to relate the functionally important transmembrane fluxes of Ca2+, K+ and Cl- to one another and to changes in membrane potential and intracellular pH to determine mechanisms of control of these events and the consequences of their occurrence, and; 2. to examine the role of cAMP and inositol polyphosphates (IP) in lytic function and to relate changes in the intracellular concentrations of these second messengers to ion fluxes as well as to determine the possible role of cAMP in control of protein phosphorylation and antigen induced morphologic changes in CTL. These studies will use fluorescence spectroscopy, flow cytofluorimetry, and high resolution cinemicrography to document the involvement of ionic species in CTL mediated lysis and to relate these events to morphological changes which accompany the lytic process. Additionally, various pharmacological agents will be used to inhibit cytolysis by distinct mechanisms. Phosphoprotein analysis, measurement of intracellular cAMP and IP as well as analysis of G protein involvement will be conducted to relate various inhibitory manipulations to known regulatory pathways. These observations will be synthesized into a pathway leading from target cell binding to cytolysis.

该提议的长期目标是实现全面了解信号转导和致命命中的机制在细胞溶解T淋巴细胞（CTL）中递送。 CTL是最终效应器免疫反应的细胞臂中的细胞，并参与宿主防御肿瘤和病毒疾病以及移植的拒绝和移植-VS宿主疾病。因此，这些细胞是激活以产生这些影响是基本的生物医学重要性。该提案的具体目的是：1。 Ca2+，K+和Cl-的重要跨膜通量，彼此之间膜电位和细胞内pH的变化以确定机制控制这些事件及其发生的后果，以及； 2。检查cAMP和肌醇多磷酸盐（IP）在裂解中的作用功能并关联这些细胞内浓度的变化第二使者到离子通量以及确定可能的作用控制蛋白质磷酸化和抗原诱导的cAMP CTL的形态变化。这些研究将使用荧光光谱，流式细胞荧光法和高分辨率摄影记录离子物种参与CTL介导的裂解和将这些事件与伴随裂解的形态变化联系起来过程。此外，将使用各种药理学剂通过不同的机制抑制细胞解。磷蛋白分析，细胞内营地和IP的测量以及G蛋白的分析将进行参与，以将各种抑制作用与已知的监管途径。这些观察结果将合成从靶细胞结合到细胞溶解的途径。