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Contenido archivado el 2024-04-30

Identification of tyrosine kinases involved in phagocytosis and phagosome-lysosome fusion

Objetivo



Research objectives, and content
Phagocytosis and phagosome-lysosome (PLF) fusion are key events in killing of microorganisms. Certain bacteria, such as Mycobacterium tubercolosis (Mtb) and Chlamydia trachomatis (Ct), can survive within phagosomes which do not fuse with lysosomes. Recent evidence suggested that protein tyrosine phosphorylation may represent a key signal in mediating phagocytosis and PLF. My project is to address several topics related to the role played by tyrosine kinases in phagocytosis of pathogens and in PLF. These topics can be summarized as follows. 1. Activation of tyrosine kinase activities during phagocytosis of complement- or IgG-opsonized particles, and Mtb and Ct in human neutrophils and monocytes. This topic will be addressed analyzing activity of distinct tyrosine kinases in phagocytes challenged with injecstible particles. Kinases analyzed will be Src-family tyrosine kinases (Lyn, Fgr, Hck) and p72syk. The specific aim of this line of experimentation is to identify kinases involved in phagocytosis and possibly PLF 2. Redistribution of tyrosine kinases, tyrosine phosphorylated and cytoskeletal proteins and annexins to the phagosome surface following phagocytosis of complement- or IgG-opsonized particles, and Mtb and Ct. This topic will be addressed by fluorescence microscopy analysis. The specific aim of this line of experimentation is to define proteins redistributed to the phagosomal membrane and possible alterations of this redistribution during phagocytosis of Mtb and Ct. 3. Analysis of phagocytosis, PLF and bacterial growth and survival in neutrophils and macrophages isolated from mice with the selective deficiency of the Src-family tyrosine kinases Fgr, Hck and Lyn. This topic will be addressed by using phagocytes from mice with targeted disruption of genes eneoding Sre-family tyrosine kinases. The specific aim of this line of experimentation is to provide a formal proof of the possible role of Src family kinases in phagocytosis, reorganization of the cytoskeleton and PLF during phagocytosis of Mtb and Ct.
Training content (objective, benefit and expected impact)
One of the main topic of the research activity of the laboratory in which I have been working has been elucidation of transmembrane signaling involved in phagocytosis and PLF. The expertise I acquired matches perfectly with the interest of the laboratory of Dr. Giorgio Berton on the role of tyrosine kinases in signaling in phagocytic cells. The benefit of combining these two different expertise in one well defined research project is to elucidate new aspects of regulation of phagocytosis and PLF. I expect to acquire new methodologies to study transmembrane signaling and exploit these to push a step forward the understanding of mechanism of regulation of phagocytosis and PLF.

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Coordinador

UNIVERSITA DEGLI STUDI DI VERONA
Aportación de la UE
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Dirección
Strada le Grazie 8
37134 VERONA
Italia

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Participantes (1)