CORE--CYTOKINE/IMMUNOLOGY FACILITY

细胞因子/免疫学核心的具体目标是为该计划项目资助的研究人员分析复杂的小鼠生物液体和组织中的细胞因子和粘附分子。此外，该核心将提供过继转移以及骨髓嵌合体实验。计划项目中提出的四个项目侧重于涉及细胞因子作用的研究，计划项目中提出的炎症项目侧重于涉及细胞因子和炎症细胞在实验克罗恩病中作用的研究。因此，将需要细胞因子测量的定量和标准化以及过继转移和骨髓嵌合体实验。将在体外实验和组织提取物中通过多种技术和多个水平定量细胞因子。具体而言，将通过北方印迹分析、RNA酶保护试验以及半定量逆转录聚合酶链反应评估mRNA水平;将根据RNA的可用性和量选择试验。此外，将通过ELISA和Western印迹分析测量蛋白质水平。将使用各种T细胞亚群实现连续转移实验，并将以集中方式对所有项目进行骨髓嵌合体，以减少实验之间的差异性、结果的标准化并减少这些实验中使用的小鼠总数。这些方法是由主任和共同主任的实验室制定的，目前正在使用。此外，该核心还将开发新的检测方法，以根据项目2和3以及其他两个项目之间的相互作用识别新的细胞因子和/或粘附分子。因此，细胞因子/免疫学核心的总体目标是为细胞因子和粘附分子的定量分析以及细胞免疫学研究提供专业知识和技术支持，并实施最先进的方法。

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.