whether the virus subverts the autophagy process in human monocytes and macrophages, the principal cell type involved in the pathogenesis of dengue. To undertake the work, primary human macrophages and monocytes will be used, as well as a monocytic cell line U937, a cell line which is known to closely mimic the responses of normal primary human monocytes. In addition, further analysis of the autophagy induction in HepG2 cells will be undertaken as this is a cell line in which the laboratory has considerable experience. In particular the project will seek to define the relationship between the dengue replication complex and autophagic membranes. This will be achieved by co-localization of dengue markers (NS1, E and double stranded RNA) and specific markers of the autophagic machinery. We will also attempt to document the presence of the translation machinery for dengue by co-localization of ribosomal specific markers. The work in HepG2 cells will also seek to define more closely the relationship between autophagic vacuole maturation and the dengue virus. In particular the relationship between dengue markers and markers of the late stage (autophagolysosomes) autophagy process will be investigated.
