Abstract Title
Role of E1 Gene in Ad14p1 Pathogenesis
Additional Funding Sources
The project described was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant No. P20GM103408.
Abstract
Adenovirus 14p1 (Ad14p1) is an emergent variant of Adenovirus 14 (Ad14) which has had an increased pathogenicity in the past outbreaks. Infection with Ad14p1 can result in acute respiratory distress syndrome (ARDS), an inflammatory disease that is controlled by macrophages. Reduced expression of the Ad gene E1B 20K in Ad14p1 infected cells renders the infected cells unable to repress inflammatory responses of macrophages. Our hypothesis is that mutations within the E1B promoter in Ad14p1 is responsible for the increased pathogenesis. The objective of my research is to create tools to test this hypothesis. To test if the E1 region of Ad14p1 is the viral region that controls pathogenesis, we have created chimeric viruses by swapping the E1 regions of Ad14 and Ad14p1. To determine if mutations in the Ad14p1 E1B promoter result in reduced expression of E1B 20K, we have cloned the E1B promoters in a promoter-less Luciferase reporter vector and using site directed mutagenesis we are screening each mutation and combination of mutations on the efficiency of the Ad14p1 E1B promoter. The optimized Ad14p1 E1B promoter can then be cloned into Ad14p1 virus for pathogenesis studies in our Syrian hamster model.
Role of E1 Gene in Ad14p1 Pathogenesis
Adenovirus 14p1 (Ad14p1) is an emergent variant of Adenovirus 14 (Ad14) which has had an increased pathogenicity in the past outbreaks. Infection with Ad14p1 can result in acute respiratory distress syndrome (ARDS), an inflammatory disease that is controlled by macrophages. Reduced expression of the Ad gene E1B 20K in Ad14p1 infected cells renders the infected cells unable to repress inflammatory responses of macrophages. Our hypothesis is that mutations within the E1B promoter in Ad14p1 is responsible for the increased pathogenesis. The objective of my research is to create tools to test this hypothesis. To test if the E1 region of Ad14p1 is the viral region that controls pathogenesis, we have created chimeric viruses by swapping the E1 regions of Ad14 and Ad14p1. To determine if mutations in the Ad14p1 E1B promoter result in reduced expression of E1B 20K, we have cloned the E1B promoters in a promoter-less Luciferase reporter vector and using site directed mutagenesis we are screening each mutation and combination of mutations on the efficiency of the Ad14p1 E1B promoter. The optimized Ad14p1 E1B promoter can then be cloned into Ad14p1 virus for pathogenesis studies in our Syrian hamster model.