CORE--CYTOKINE/IMMUNOLOGY FACILITY

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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.