EFFECTS OF CULTURE CONDITIONS ON INTERFERON-gamma PRODUCTION IN WHOLE BLOOD CULTURES FROM MYCOBACTERIUM BOWS-INFECTED CATTLE

摘要

The cell-mediated immune response to antigens of M.bovis is currently used in the form of the delayed hypersensitivity test in skin or, in vitro, by the production of IFN-gamma by T-cells in whole blood culture. The analysis of various parameters of the IFN-gamma test is done in view of potential simplifications in order to streamline the assay procedure. Here we show that the culture temperature needs to be 33°C or higher and that there is no need to use incubation in the presence of C02. Furthermore, various plate formats are shown to be feasible. The produced IFN-y was stable at 4°C for 28 days as well as after repeated freeze-thaw cycles. It is therefore conceivable to start the stimulation part out in the field allowing for a more efficient testing procedure.