Detection of specific T cell reactivity in sheep infected with Mycobacterium avium subspecies silvaticum and paratuberculosis using two defined mycobacterial antigens.

A 30 kDa antigen (P30) from Mycobacterium avium ssp. paratuberculosis (M. a. paratuberculosis) and a 40 kDa (P40) antigen from Mycobacterium avium ssp. silvaticum (M. a. silvaticum) were employed in two different assays to measure the cell-mediated immune reactivity of ovine peripheral blood lymphocytes. In lymphocyte stimulation assays, proliferative responses to the P30 were observed only with lymphocytes from sheep inoculated with live M. a. paratuberculosis or M. a. silvaticum. Although this antigen was not subspecies-specific it differentiated between animals given live organisms and those inoculated with an inactive lysate. The P40 protein from M. a. silvaticum showed subspecies specificity by eliciting in vitro responses only with lymphocytes derived from sheep inoculated with live M. a. silvaticum. Similar results were obtained using an interferon-gamma release assay which proved to be a more rapid and sensitive system.