CORE--CYTOKINE/IMMUNOLOGY FACILITY

细胞因子/免疫学核心的具体目标是为本计划项目资助的研究人员分析复杂小鼠生物体液和组织中的细胞因子和粘附分子。此外，该核心将提供过继性转移以及骨髓嵌合体实验。本项目拟建的4个项目主要研究细胞因子和炎症的作用。本项目拟建的项目主要研究细胞因子和炎症细胞在实验性克罗恩病中的作用。因此，细胞因子测量的量化和标准化以及过继移植和骨髓嵌合体实验将是必要的。细胞因子将在体外实验和组织提取中通过多种技术和多个水平进行量化。具体来说，mRNA水平将通过Northern blot分析、RNAse保护实验以及半定量逆转录聚合酶链反应来评估；将根据RNA的可用性和数量选择检测方法。此外，蛋白质水平将通过ELISA和Western blot分析测定。过继性转移实验将使用各种T细胞亚群来实现，骨髓嵌合体将在所有项目中以集中的方式进行，以减少实验之间的可变性，结果的标准化和减少在这些实验中使用的小鼠总数。这些方法是由主任和副主任的实验室开发的，目前正在使用。此外，该中心还将根据项目2和项目3以及其他两个项目之间相互作用产生的新细胞因子和/或粘附分子的鉴定需求，开发新的检测方法。因此，细胞因子/免疫学核心的总体目标是提供专业知识、技术支持和实施最先进的方法，用于细胞因子和粘附分子的定量分析以及细胞免疫学研究。

The specific goals of the Cytokine/Immunology Core are to carry out the analysis of cytokine and adhesion molecules in complex murine biological fluids and tissues for investigators in this Program Project Grant. In addition, this Core will provide adoptive transfer as well as bone marrow chimera experiments. The four projects proposed in the Program Project focus on research involving the role of cytokines and inflammatory projects proposed in the Program Project focus on research involving the role of cytokines and inflammatory cells in experiment Crohn's disease. Therefore, quantification and standardization of cytokine measurements as well as adoptive transfer and bone marrow chimera experiments will be required. Cytokine will be quantified in vitro experiments and tissue extracts by a variety of techniques and at multiple levels. Specifically, mRNA levels will be assessed by Northern blot analysis, RNAse protection assay, as well as semi-quantitative reverse transcription polymerase chain reaction; assay will be chosen based on the availability and amount of RNA. In addition, protein levels will be measured by ELISA and Western blot analysis. Adoptive transfer experiments will be achieved using a variety of T cell subsets, and bone marrow chimeras will be performed in a centralized fashion for all projects in order to decrease variability between experiments, standardization of results and decrease total number of mice used in these experiments. These methods were developed by the Director and co- Director's laboratories and are currently in use. In addition, the core will also develop new assays will be developed as the need emerges from the identification of new cytokines and/or adhesion molecules from the interactions between Projects 2 and 3 as well as the other two projects. The overall objectives of the Cytokine/Immunology Core therefore are to provide expertise as well as technical support and implementation of state-of-the-art methodologies for the quantitative analysis of cytokines and adhesion molecules as well as cellular immunological studies.