PHYSICIAN SCIENTIST AWARD

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Monocyte stimulation resulting in interleukin-1 (IL-1) production can be accomplished with many different compounds. Yet little insight into the negative regulation of IL-1 production is available. Therefore, this research proposal is a two-phase project aimed at elucidating control of production of this monokine. In phase one, we will begin to identify in monocytes 1) time course of IL-1 protein and gene expression, 2) the effects of different stimuli on differential IL-1 gene expression, 3) pharmacological inhibition of IL-1 protein or gene expression, 4) the role of arachidonic acid metabolites in IL-1 protein or gene expression, 5) lymphocytes and lymphocyte products' place in regulation of IL-1 protein or gene expression. Experiments will be performed on peripheral blood monocytes stimulated with LPS or other IL-1 inducers. Inhibition experiments will evaluate commonly used immunomodulatory drugs (dexamethasone and cyclosporin A), cyclooxygenase (indomethacin) and lipoxygenase (U60) inhibition of arachidonic metabolism. Analysis of IL-1 protein expression will require use of standard tissue culture techniques for monocyte isolation, mouse thymocyte bioassays for IL-1 functional quantification and Northern analysis of total cellular RNA using a synthetic oligonucleotide directed against IL-1 mRNA. In phase two, application of immunological control mechanisms will be applied to alveolar macrophage and sarcoidosis, where we hypothesize that abnormal IL-1 regulation may lead to permanent disability. We will identify in normal alveolar macrophages and in sarcoidosis 1) time course of IL-1 protein and gene expression, 2) the effects of different stimuli on differential IL-1 gene expression, 3) pharmacological inhibition of IL-1 protein and gene expression, 4) the role of normal lymphocytes, their products and those in sarcoidosis in regulating IL-1 protein and gene expression. These studies will be accomplished through use of standard mouse thymocyte assays and Northern analysis of IL-1 RNA. With pharmacologically relevant inhibitors studied in detail, new therapeutic options will then become available for immunologically mediated lung disease, such as sarcoidosis. Regulation of IL-1 production may be very important in altering prognosis and functional impairment in these diseases.