Scuticociliate proteinases may modulate turbot immune response by inducing apoptosis in pronephric leucocytes
| Autor: | Rosario Castro, M. L. Sanmartín, Jesús Lamas, A. Paramá, M. T. Santamarina, José Leiro |
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| Rok vydání: | 2007 |
| Předmět: |
Programmed cell death
Cell Survival Immunoblotting Apoptosis Ciliophora Infections Caspase 3 DNA Fragmentation Biology Kidney Fish Diseases chemistry.chemical_compound Affinity chromatography Leukocytes Animals DAPI Ciliophora Protozoan Infections Animal Cell Nucleus Electrophoresis Agar Gel Innate immune system DNA Protozoan Molecular biology Cysteine Endopeptidases Infectious Diseases chemistry Flatfishes DNA fragmentation Electrophoresis Polyacrylamide Gel Parasitology Peptide Hydrolases Cysteine |
| Zdroj: | International Journal for Parasitology. 37:87-95 |
| ISSN: | 0020-7519 |
| DOI: | 10.1016/j.ijpara.2006.08.013 |
| Popis: | The role of proteinases of the histiophagous ciliate Philasterides dicentrarchi, purified by affinity chromatography in bacitracin-Sepharose, on apoptosis (programmed cell death) of turbot pronephric leucocytes (PL) was investigated. The results showed that more than 90% of proteinases purified by bacitracin-Sepharose were cysteine proteinases, which lacked significant caspase-3-like activity and generated three main gelatinolytic bands of molecular weights 36, 45 and 77 kDa as determined by gelatine-SDS-PAGE and immunoblot. Viability of PL cells after 24 h stimulation with P. dicentrarchi cysteine proteinases did not differ from that of non-stimulated cells. Apoptosis was confirmed by: (i) caspase activity, (ii) DNA fragmentation, and (iii) nucleus fragmentation. The caspase-3-like activity in PL incubated for 4h in the presence of 125, 250 and 500 microg/ml of proteinases increased in a dose-dependent fashion. The PL DNA was fragmented following 24-h exposure to P. dicentrarchi cysteine proteinases and characteristic DNA ladders consisting of multimers of approximately 180-200 pb were produced. Morphological changes, such as chromatin condensation and nucleus fragmentation, were observed under fluorescence microscopy after DAPI staining of the PL cells incubated with cysteine proteinase-incubated for 24 h. The results suggest that the pathogenic scuticociliate P. dicentrarchi may induce host leucocyte programmed cell death via the production of cysteine proteinases, as a mechanism of pathogenesis and evasion of the turbot innate immune response. |
| Databáze: | OpenAIRE |
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