Tyrosine kinase-mediated cell signalling in the activation of Mytilus hemocytes: possible role of STAT-like proteins.
In bivalve molluscs, cell-mediated immunity is carried out by circulating hemocytes, resembling the monocyte/macrophage lineage of vertebrates, that can kill the microbes through phagocytosis and various cytotoxic reactions. Previous data demonstrated that activation of MAPKs (Mitogen Activated Protein Kinases) is involved in the response of mussel hemocytes (Mytilus galloprovincialis Lam.) to bacterial challenge. In this work, the possibility that modulation of components of tyrosine kinase-mediated cell signalling may participate in the activation of mussel hemocytes was investigated. Cell pre-treatment with the macrophage activator IFN gamma significantly increased the bactericidal activity of mussel hemocytes towards E. coli. Human recombinant IFN gamma stimulated tyrosine phosphorylation of different members of STAT-like proteins (Signal Transducers and Activators of Transcription), as evaluated by Western blotting of hemocyte protein extracts with specific anti-phospho-STAT antibodies. A similar increase in phosphorylation of immunoreactive STATs was observed in hemocytes incubated with E. coli, this indicating that tyrosine phosphorylation of STAT-like members represents a physiological step in hemocyte activation. IFN gamma lead to persistent phosphorylation of immunoreactive STAT1, a transcription factor that plays a critical role in innate immunity towards Gram negative bacteria in mammalian systems; moreover, hemocyte pretreatment with IFN gamma significantly increased bacteria-induced STAT1 phosphorylation, whereas IFN alpha did not. IFN gamma also transiently affected the phosphorylation state of different MAPKs. The extent and time course of MAPK phosphorylation induced by IFN gamma were distinct from those elicited by either IFN alpha or bacterial challenge. Overall, the results indicate that the hemocyte function can be modulated by heterologous cytokines and bacterial signals that act in concert through tyrosine kinase-mediated transduction pathways converging on STAT- and MAPK-like members.
Canesi
Laura
L
Istituto di Scienze Fisiologiche, Università di Urbino, Loc. Crocicchia, Urbino, 61029 (PU), Italy. canesi@uniurb.it
Betti
Michele
M
Ciacci
Caterina
C
Citterio
Barbara
B
Pruzzo
Carla
C
Gallo
Gabriella
G
D000818:Animals; D049872:Bivalvia / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000254:growth & development; D006434:Hemocytes / Q000187:drug effects / Q000382:microbiology / Q000502:physiology*; D007371:Interferon-gamma / Q000494:pharmacology; D008894:Milk Proteins / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D050794:STAT1 Transcription Factor; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D015398:Signal Transduction / Q000502:physiology*; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D004268:DNA-Binding Proteins; D008894:Milk Proteins; D050794:STAT1 Transcription Factor; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D015534:Trans-Activators; D014443:Tyrosine; D007371:Interferon-gamma; D011505:Protein-Tyrosine Kinases; D020928:Mitogen-Activated Protein Kinases