PATHOGENESIS AND TREATMENT OF MULTIPLE SCLEROSIS

The proposed program extends and develops further investigations into immunopathogenesis of the demyelinative disorders and the structural and functional effects of those factors. Project I- will examine the effects of lymphokines, T-cells and serum on the integrity of myelin in organotypic cultures as determined by immunocytochemical and structural observations. Co-cultures of CNS fragments and endothelial cells will be established in an in vitro counterpart of our previous studies of the blood-brain barrier. Project II- will involve the expression, cellular localization and immunopathologic effects of products of the immune system that may act in immune-mediated demyelination by means of ultrastructural and immunocyte chemical techniques applied to cultured tissues affected by lymphocytes, T-cell lines and cytokines. Project III -will involve continuations of analyses of passively transferred EAE using new markers (T-200, PMN leucocytes, interleukin-2) and transferrin receptors, interleukins, interferons and tumor necrosis factor, adhesion molecules, and receptors for high endothelial venules, organotypic cultures will be used to examine the roles of various growth factors, and adhesion molecules during demyelination and remyelination. ProjectIV- will examine epidural visual evoked potentials, direct intracranial recordings of unit activity within tracts and cortex will be combined with monocular injection of cytokines and antibodies against specific CNS antigens. Project V- Will explore in greater detail the role of cytokines in inducing changes in the cerebral vasculature leading to inflammation of the CNS. Biological assays and molecular biological techniques will be used. Project VI- will assess GFAP mRNA levels as they may parallel GFA protein by in situ hybridization analysis in sections of Lewis rat cord and brain and in primary rat astrocyte cultures and human astrocytoma cell line HTB-17. Core A- will supply organtoypic cultures for this program and, in addition, will establish cultures of endothelial cells and CNS tissue.