benzyloxycarbonylvalyl-alanyl-aspartyl-fluoromethyl-ketone and Paragonimiasis

Eosinophils are important cells in host immunity to infections by parasitic worms. Tissue-invasive helminthic parasites have been known to induce apoptosis of immune cells for their successful establishment in vivo. We have previously found that high doses of excretory-secreted products (ESP) secreted by lung fluke Paragonimus westermani newly excysted metacercariae (PwNEM), which cause pulmonary or extrapulmonary paragonimiasis in human beings, accelerate eosinophil cell death. However, little is known about the mechanism of eosinophil apoptosis induced by the Paragonimus-derived products.. We examined involvement of caspase 3 activation in eosinophil cell death induced by ESP produced by PwNEM.. Eosinophils were isolated from the peripheral blood of healthy donors by CD16-negative immunomagnetic selection. We examined the inhibitory effect of pan-caspase inhibitor on ESP-triggered phosphatidylserine (PS) externalization on the outer surface of eosinophils and intracellular activation of caspase 3 in cell lysates treated with the ESP.. When ESP secreted by PwNEM were incubated for up to 3 h with eosinophils, they increased surface exposure of PS on eosinophils in a time- and dose-dependent manner. This proapoptotic effect of the ESP on eosinophils was significantly inhibited by pretreatment of cells with pan-caspase inhibitor z-VAD-fmk, and was completely abolished by heat inactivation of ESP at 56 degrees C. The activated forms of caspase 3 were also clearly detected in eosinophils incubated with ESP. Moreover, ESP potently inhibited prolonged survival of eosinophils induced by cytokines such as IL-5, IL-3, and GM-CSF.. These results suggest that ESP secreted by PwNEM contain biological active factors causing caspase-3-mediated apoptosis of human eosinophils, thereby enabling the larvae to evade and to subvert the tackle by eosinophils during the early phase of the infection.

Topics: Amino Acid Chloromethyl Ketones; Animals; Antigens, Helminth; Apoptosis; Caspase 3; Caspase Inhibitors; Caspases; Cell Survival; Cysteine Proteinase Inhibitors; Eosinophils; Flow Cytometry; Granulocyte-Macrophage Colony-Stimulating Factor; Hot Temperature; Humans; Interleukin-3; Interleukin-5; Kinetics; Paragonimiasis; Paragonimus; Phosphatidylserines