Metabolism of the TMA group of antigens during the growth cycle of mycobacteria

The TMA (thermostable macromolecular antigens) group includes A60 of Mycobacterium tuberculosis and A7 of M. leprae, active components of tuberculin and lepromin. We have previously described the purification and composition of A60, and its ability to elicit immune reactions of humoral and cellular type. In the present work, the intracellular and extracellular distribution and composition of A60 have been traced, as a function of the replication cycle, in static surface cultures of M. bovis. In exponentially-growing mycobacteria, most A60 was present in the cytoplasm and had a high protein/polysaccharide ratio: this ratio, as well as the level of cytoplasmic A60, decreased after cessation of cell proliferation. The A60 fraction located within the cell wall increased during the stationary phase, but its protein/polysaccharide ratio underwent minor changes. A release of cellular polypeptides and polysaccharides into the extracellular fluid occurred during the declining and lysing phases: a fraction of it was represented by A60. This explains the practice of old tuberculin preparation by autoclaving filtrates of autolysed mycobacterial cultures. The pattern of an A60-like antigen in shaken homogeneous cultures of M. smegmatis was similar (most antigen present in cytoplasm during growth, increase of the wall fraction in stationary phase, and extracellular release during the declining phase)