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Comparative structure-function analysis of the dengue virus envelope protein in complex with relevant ligands from the host


We propose to solve the crystal structures of the dengue virus (DV) envelope (E) protein from different serotypes in complex with immunologically relevant ligands from the human host.

These will be:
- Dendritic cell-specific ICAM-3 grabbing non-integrin (DC -SIGN). This molecule has been shown to be essential for entry of mosquito-grown virus into human dendritic cells.
- Fab fragments from antibodies capable of neutralising one or more DV serotypes.

In both cases we will be especially interested in serotype de pendent variations and their functional consequences, and will use a comprehensive range of DV field strains isolated from dengue fever patients in Venezuela in an extensive comparative study.

Analysis of the crystal structures will be performed in the con text of binding studies of the ligands with both intact virus and purified E-protein from the corresponding strains. In the case of DC-SIGN the results will be correlated with epidemiological data on the relative virulence of the different serotypes.

This study will elucidate the details of the key molecular interactions at the host cell surface, which occur during the primary DV infection at the site of the mosquito bite. Furthermore, it will reveal the specific antigenic determinants of the E-protein and their serotype dependence, which will be extremely valuable for the development of a safe DV vaccine.

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