John Anthony Fraga

John Anthony Fraga

PI: James Crowe, MD , Vanderbilt Vaccine Center, Departments of Pathology, Microbiology and Immunology & Pediatrics

Isolation and characterization of human monoclonal antibodies for novel anti-viral treatments of Ross River and rabies viruses

Ross River virus (RRV) is a mosquito-transmitted viral disease that is endemic to Australia and the South Pacific region, with 5,000-8,000 cases occurring every year. RRV belongs to the Alphavirus genus and is characterized by symptoms including polyarthritis, fever, and rashes. Rabies virus (RABV) is a worldwide viral disease belonging to the Rhabdoviridae family that causes acute encephalitis and has the highest human case-fatality proportion of all conventional infectious diseases. Currently no treatment or licensed vaccine is available for RRV, and methods of treatment for RABV have yet to incorporate monoclonal antibodies that can broadly neutralize all RABV strains. In this study, we will isolate, expand, and characterize both RRV and RABV human monoclonal antibodies (mAbs) from the blood of a single donor who has acquired immunity to both viruses. We will transform B cells from this donor with Epstein-Barr virus and fuse selected EBV B cell lines with a myeloma partner to create human hybridomas that produce human mAbs with high affinity and functional activity. We also will perform neutralization assays of current RRV mAbs against other alphaviruses, such as Chikungunya, Sagiyama, and Getah viruses, to test for potency and cross-reactivity. Furthermore, we will attempt to produce escape mutant viruses for RRV neutralizing antibodies in order to understand what sites of the RRV genome are targeted by these antibodies. Discovered antibodies from our findings could lead to possible treatment/prevention of Ross River disease and more broadly neutralizing methods of treatment for RABV.