Understanding the Role of Envelope Protein of Tick-borne Flaviviruses in Vector Specificity
Restricted (Penn State Only)
Author:
Wang, Anqi
Area of Honors:
Biochemistry and Molecular Biology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Joyce Jose, Thesis Supervisor Song Tan, Thesis Honors Advisor
Keywords:
Virology biochemistry flavivirus chimera
Abstract:
Flaviviruses are arthropod-borne viruses infecting humans and causing debilitating diseases. For tick-borne and mosquito-borne flaviviruses, vector specificity and tropism are poorly understood. We aimed to investigate the role of Powassan virus envelope (E) protein, which is crucial for viral entry and tick vector specificity. We successfully constructed cDNA clones and generated chimeric yellow fever-Powassan (YFV-DTV and YFV-POWV) viruses, which could infect mosquitoes and various mammalian cell lines. By performing growth kinetics analysis, the infectivity of chimeric viruses (YFV-DTV and YFV-POWV) was assessed and compared across multiple cell lines with wild-type yellow fever virus (YFV) as the control. Furthermore, we performed virus entry assays with wild-type YFV and chimeric viruses (YFV-DTV and YFV- POWV) in cell lines such as A549 (human lung carcinoma cell line), C6/36 (mosquito cell line), and BHK-15 (baby hamster kidney cell line). Our assays revealed differences in the efficiency of viral entry of YFV and chimeric viruses (YFV-DTV and YFV-POWV) due to variance in prM and E proteins. Furthermore, we successfully generated a BHK-15 cell line that stably produces POW-VLP, which is a promising vaccine candidate. In conclusion, we have gathered evidence that structural proteins play important roles in the tropism of POWV. Lastly, the chimeric viruses we created will serve as vaccine candidates to treat POWV infection.
