The causal agent of heartwater disease of domestic ruminants, Cowdria ruminantium, can, with difficulty, be isolated and passaged in lines of bovine endothelial cells grown in the presence of the Glasgow modification of Eagle's minimal essential medium. However, when Leibovitz's L-15 medium supplemented with 0.45% glucose at pH 6.0-6.5 is used as maintenance medium for these cells, isolation and serial passage may routinely be achieved.
Conconavalin A-stimulated bovine T-cell supernatants inhibited the growth of Cowdria ruminantium in bovine endothelial cells in vitro but did not affect their entry. This finding represents one mechanism by which T cells may control C. ruminantium multiplication and hence affect the severity of disease.
In an earlier study we demonstrated that Concanavalin-A stimulated bovine T cell supernatants inhibited the growth of Cowdria ruminantium in bovine endothelial cells in vitro. An investigation was conducted to identify the cytokines which were responsible for this growth inhibition. Addition of antiserum against bovine interferon gamma (IFN gamma) reproducibly neutralized the inhibitory effect of the T cell supernatants, whereas addition of antisera against bovine tumor necrosis factor alpha (TNF alpha) had no effect. The inhibitory effect of IFN gamma on C. ruminantium growth was not mediated by the production of nitric oxide as there was no detectable difference in nitric oxide levels in cultures that were supplemented with T cell supernatants compared with those that were not. The IFN gamma mediated anti-C. ruminantium effect highlights the importance of cell mediated immune responses in control of these infections and in particular incriminates the protective role of T cells, or cells that secrete IFN gamma.
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