The processing and presentation of mycobacterial antigens by human monocytes

The processing and presentation of whole irradiated Mycobacterium tuberculosis (Mtb gamma) and its purified protein derivative (PPD) by the peripheral blood monocytes from healthy Bacillus Calmette Guérin (BCG)-vaccinated individuals was investigated. To study processing and presentation as events distinct from T cell recognition and proliferation, monocytes were pulsed with antigens for varying time intervals and fixed. The kinetics of presentation indicate that up to 2 h was required for effective presentation of PPD and 2-4 h for Mtb gamma, and that the ability to activate T cells declined as the time interval for which pulsing occurred was increased, so that responses were abolished by 8-10 h. Prefixed monocytes could not present Mtb gamma and PPD to T cells indicating that processing was an essential requisite. Lysosomotropic agents chloroquine, monensin, and leupeptin inhibited the presentation of these antigens suggesting the role of lysosomes/endosomes in processing. Furthermore, monocytes incubated with optimal concentration of antigens for different lengths of time released determinants which were still antigenic but circumvented the need for any further processing. Addition of nonprimed syngeneic monocytes, both untreated or paraformaldehyde fixed to cells which had been pulsed and fixed, restored the responses even at the later time periods when responses were not detected. This second interaction of the monocyte with T cells was not major histocompatibility complex restricted in that the addition of monocytes from another donor was equally effective.