Echinococcus multilocularis metacestode metabolites contain a cysteine protease that digests eotaxin, a CC pro-inflammatory chemokine.
In many helminthic infections, eotaxin, a CC-chemokine, triggers the mobilization of eosinophils, thus, contributing to an elevated blood and periparasitic eosinophil level. Following an experimental intraperitoneal infection of C57BL6 mice with Echinococcus multilocularis metacestodes, however, we observed the absence of eosinophils in the peritoneal cavity and a low number of such cells in the blood of infected animals. Therefore, we carried out an explorative study to address the question why eosinophilia did not occur especially in the peritoneal cavity of such secondarily AE-infected mice. In an in vitro assay, we showed that metacestode antigens (in vitro generated vesicle fluid and E/S products) were able to proteolytically digest eotaxin. This effect was confirmed with semiquantitative Western blotting, which demonstrated a decreasing intensity of remaining eotaxin signals. Proteolysis of eotaxin was, thus, dose-dependent and proportional to the time of incubation with the metacestode antigens. Using appropriate inhibitors, the respective protease was identified as a cysteine protease, which required the presence of Ca(++) as co-enzyme. A chromatographic fractionation procedure by successive separation of VF molecules using a superpose column and subsequently a MonoQ column mounted on an FPLC system allowed to yield a fraction, referred to us as fraction 6; containing the enriched cysteine protease, this fraction will be used for further molecular studies. Eotaxin inactivation by VF and E/S products may contribute to explain the absence of eosinophils within the peritoneal cavity of AE-secondary infected mice. Absent eosinophils, thus, may be a part of a series of events that maintain a low level of inflammation displayed within the peritoneal cavity of experimentally infected mice.
Mejri
N
N
Institute of Parasitology, University of Berne, 3012 Bern, Switzerland.
Gottstein
B
B
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D054413:Chemokine CCL11 / Q000378:metabolism*; D002853:Chromatography, Liquid; D003067:Coenzymes; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D004195:Disease Models, Animal; D004443:Echinococcosis / Q000276:immunology*; D048210:Echinococcus multilocularis / Q000201:enzymology* / Q000276:immunology*; D004804:Eosinophils / Q000276:immunology*; D005260:Female; D007814:Larva / Q000201:enzymology / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D013997:Time Factors
D002413:Cations, Divalent; D054413:Chemokine CCL11; D003067:Coenzymes; D003546:Cysteine Endopeptidases; D002118:Calcium