Identification of bovine T cell stimulatory antigens using a cosmid library of Mycobacterium bovis in Mycobacterium smegmatis

Abstract

Culture filtrates derived from a Mycobacterium bovis cosmid library in Mycobacterium smegmatis were screened for T cell antigens. Recognition and reactivity were measured by the levels of lymphocyte proliferation and the levels of gamma interferon (IFN-γ) produced when the culture filtrates were incubated with peripheral blood mononuclear cells (PBMC) taken from cattle immunised with M. bovis BCG. The screening system was optimised to distinguish between M. bovis secreted antigens and normal M. smegmatis secreted proteins. From ten culture filtrates screened, two were identified that induced lymphocyte proliferation and IFN-γ production. Analysis of the DNA inserts from the recombinant cosmids suggest that they may code for different proteins. The results demonstrate that screening recombinant M. smegmatis culture filtrates can be used to identify M. bovis T cell antigens that are recognised by immunised cattle. These antigens may be important for the develoument of vaccines with protective ability against bovine tuberculosis.