IN VIVO ROLE OF CRP IN TRANSGENIC MICE

本计划的总体目标是定义C的作用 反应蛋白（CRP）在宿主防御机制中的作用。 已经 假设，主要基于体外研究，CRP功能 在炎症刺激后的早期免疫前阶段。 一 CRP在体内炎症反应过程中的明确作用 尚未成立。 在上一个融资期， 研究者已经产生了表达兔CRP的转基因小鼠， 金属硫蛋白（MTT）或PEPCK启动子， CRP的表达不依赖于炎症刺激。 这些转基因 动物已经被用于三种小鼠炎症模型中， 包括LPS和PAF的全身效应，肺泡炎模型， 炎症和单关节关节炎模型。 在所有三 系统表达CRP的转基因小鼠已经显示出可重复的 影响。 目前的建议是延长研究 CRP作为炎症参与者的机制， 假设是CRP的作用取决于其能力， 结合磷酸胆碱（PC），CRP可影响细胞因子的表达， 粘附分子，以及对抗原诱导的粘附作用 关节炎是通过抑制T细胞活化介导的， 免疫过程的传入臂 这些假设将通过以下方式进行检验： 1)产生表达突变CRP的转基因小鼠， 设计为不能与PC结合; 2）测量CRP的作用 对单核细胞、脾细胞和巨噬细胞中炎性细胞因子基因表达的影响， 中性粒细胞;以及3）在治疗前、治疗期间和治疗后操纵CRP水平。 在关节炎模型中免疫应答开始后， 确定CRP对T细胞活化的影响。

The overall objective of this program is to define the role of C reactive protein (CRP) in host defense mechanisms. It has been hypothesized, based largely on in vitro studies, that CRP functions during the early preimmune stages following inflammatory stimulus. A clear role for CRP during the course of inflammatory response in vivo has not been established. In the prior funding period the principal investigator has generated transgenic mice which express rabbit CRP with either a metallothionein (MTT) or PEPCK promoter which has made possible expression of CRP independent of inflammatory stimuli. These transgenic animals have been used in three mouse models of inflammation which include the systemic effects of LPS and PAF, a model of alveolar inflammation and a model of monoarticular arthritis. In all three systems CRP expressing transgenic mice have shown reproducible antiinflammatory effects. The present proposal is to extend the studies on the mechanisms of CRP as a participant in inflammation, The working hypotheses are that the effects of CRP are dependent on its ability to bind phosphocholine (PC), CRP can influence expression of cytokines and adhesion molecules, and antiinflammatory effects on antigen enduced arthritis are mediated by inhibition of T cell activation during the afferent arm of the immune process. These hypotheses will be tested by: 1) producing transgenic mice expressing a mutant CRP which has been designed to be incapable of PC binding; 2) measuring the effects of CRP on inflammatory cytokine gene expression in monocytes, splenocytes and neutrophils; and 3) manipulating the level of CRP before, during and after the initiation of the immune response in a model of arthritis to determine the effect of CRP on T cell activation.