Bovine CD4+ T-cells lines reactive with soluble and membrane antigens of Cowdria ruminantium

Cowdria-specific CD4+ T-cell lines generated from immunised cattle respond to both soluble and membrane proteins of the agent. Furthermore, the lines produced the Cowdria-inhibitory cytokine IFN-gamma in response to soluble antigens fractionated by gel filtration and FPLC. Activity eluted as a single peak around fraction 15 for all T-cell lines tested. This fraction induced the highest production of IFN-gamma by the lines and was shown by SDS-polyacrylamide gel electrophoresis and silver staining analysis to contain less than 10 different bands ranging from 22 to 32 kDa. Given their high sensitivity and specificity, these short-term CD4+ T-cell lines will be valuable tools for the identification of Cowdria antigens for incorporation in a subunit vaccine.