|Undergraduate Institution:||University of Maryland - Baltimore County
|Department:||Microbiology & Immunology|
|PI:||Arainda de Silva|
Dengue virus (DENV) is emerging as a serious disease threat. To make advancements in vaccine development and therapeutic technologies, the causal mechanisms of severe disease must be further investigated. We plan to utilize our previously-constructed DENV type 3 (DENV3) infectious clone to study how DENV genetics may contribute to viral virulence and disease outcome. The DENV3 infectious clone was constructed using a reverse genetics system and by ligating three overlapping fragments of DNA, making it easy to genetically create chimeric dengue viruses. Fragments of the infectious clone genome can be substituted with fragments from other DENV3 strains to make recombinant chimeric viruses. I will make a panel of 12 recombinant viruses containing different combinations of structural and nonstructural genes from strains linked to severe and mild epidemics. I will then use those chimeras in infection assays with human monocytic cell lines and primary human dendritic cells to determine which portion of the DENV3 genome is responsible for growth differences between strains linked to severe and mild disease. Furthermore, by using site-directed mutagenesis, I will map specific residues that are responsible for growth differences. Once I have determined the underlying genetic causes of growth differences, I will further investigate mechanistic details. The studies that I will complete with natural isolates of DENV3 and our infectious clone will provide new insight about how DENV genetics influence viral growth, pathogenesis, and disease.