| Abstrakt: |
Previous reports from this laboratory have revealed that macrophages obtained from 7-day Listeria-immune mice elicited 15 h before harvest with heat-killed homologous microorganisms were able to kill Listeria monocytogenes while resident or elicited cells were not [14, 16]. In the present study, experiments were conducted to determine if phagocytosis-associated oxidative metabolic activity participates in the enhanced destruction of Listeria by activated macrophages. Investigations into production of oxygen radicals by zymosan-stimulated macrophages revealed that Listeria-immune antigen-elicited (LIAE) cells produced significantly more superoxide and hydrogen peroxide than did resident, thioglycolate, or Listeria antigen-elicited macrophages. Additionally, the percentage of nitroblue tetrazolium (NBT) dye positive cells following exposure to zymosan was maximal in the immune-elicited population. Utilizing a luminol-dependent assay, a short-term chemiluminescent (CL) burst was noted in phagocytizing macrophages. This response was greatest in the LIAE population that exhibited a tenfold increase in peak chemiluminescence over other cell types. Prolonged in vitro culture of these cells diminishes their capacity to generate oxygen radicals. Additionally, LIAE macrophages cultured in excess of 38 h exhibited a significant decrease in zymosan-stimulated hydrogen peroxide release while the decline in superoxide generation was minimal. A substantial diminution in the Listeria-stimulated CL response was also noted during this time period. However, phagocytosis of Listeria by LIAE cells failed to induce the level of oxygen metabolites seen when zymosan was used as the particulate stimulant. In addition, scavengers of oxygen radicals were found to be relatively ineffective in reducing the killing of L monocytogenes by immunologically activated macrophages in culture. It therefore appears that toxic oxygen species do not play a major role in the heightened killing of Listeria by activated macrophages. |
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