Role of polymorphonuclear cells in Chagas' disease. I. Uptake and mechanisms of destruction of intracellular (amastigote) forms of Trypanosoma cruzi by human neutrophils.

The ability of human polymorphonuclear cells (PMN) to take up and destroy intracellular forms of Trypanosoma cruzi (AMA) was investigated as a part of our efforts to elucidate the mechanisms of clearing of these parasites from infected tissues. PMN were found to take up AMA and destroyed parasites were seen after 30 min of cell-parasite interaction. Under our experimental conditions, the rate of uptake of AMA by PMN was maximal during the first 30 min of interaction. AMA were found to be located and destroyed inside the phagolysosomal vacuoles of PMN. The parasite was never found outside these vacuoles despite electron microscopic examination of numerous preparations derived from several experiments. Intracellular destruction of AMA by PMN was visible by electron microscopy and could be monitored by measuring the release of 3H-labeled substances by PMN that had ingested radiolabeled AMA. PMN incubated after removal of unbound parasites destroyed over 90% of the ingested organisms within 3 hr and close to 99% after 12 hr. In cellfree systems, 44% of the AMA were destroyed in the presence of 10(-4) M H2O2 and all of the parasites died at 10(-3) M. Addition of lactoperoxidase and iodide resulted in 100% killing at 10(-5) M H2O2. These mechanisms appeared to be involved in the lysis of AMA by PMN since both H2O2 and peroxidase activity were demonstrated to be present in PMN vacuoles containing the parasite. Addition of NaN3, KCN (inhibitors of myeloperoxidase activity) or catalase (to decompose H2O2) caused a marked reduction in the extent of AMA killing by PMN. Xanthine oxidase was toxic for the AMA in the presence of acetaldehyde. This microbicidal activity was inhibited by catalase but not by heat-inactivated catalase or by reagents that scavenge the intermediate products of reduction of molecular oxygen, O - X 2, X OH, and 1O2. These results suggest that PMN have the potential of clearing AMA liberated in infected chagasic tissues and that parasite killing within the phagolysosomal vacuoles is mediated by myeloperoxidase activity and H2O2.