| Participant: | Lewis, Jessica |
EXPRESSION OF FLAVIVIRUS DOMAIN III PROTEINS TO ANALYZE ANTIBODY CROSS-REACTIVITY
Jessica Lewis, B.S. 1, E.I. Bovshik, MSc2, S. Zhang, M.D. 2, A.R. Lamb, B.S. 2 and D.W. C. Beasley, Ph.D. 1,2
1Departments of Pathology and 2Microbiology and Immunology, UTMB
Background: Flaviviridae is a medically important family of mosquito-borne viruses including the mosquito-borne Yellow fever, dengue, Japanese encephalitis, Saint Louis encephalitis, and West Nile viruses. Many of these viruses have overlapping geographic distribution. Although it is known that much of the antibody response induced by flavivirus infections tends to target broadly cross-reactive epitopes in the envelope (E) protein, previous studies by us and others have shown that structural domain III of E (E-III) comprises epitopes that are generally virus or sub-group specific. This suggests that the E-III portion may allow more specific serological diagnosis of flavivirus infections. Objetive: to prepare a panel of recombinant His-tagged and untagged E-III antigens representing mosquito-borne flaviviruses of global health importance and to assess, in detail, the extent of antigenic cross-reactivity that occurs following infection or vaccination of multiple species (mouse, non-human primate, human) with those viruses. Methods: In order to investigate which portion(s) of E-III are detected by specific and cross-reactive antibodies in sera from multiple species, capture of His-tagged proteins will be used to control the orientation and presentation of putative antibody binding sites, and targeted mutagenesis of individual residues identified by epitope mapping using mouse monoclonal antibodies will also be employed. The results of this work will lead to the development of improved reagents and methods for flavivirus serodiagnostics and surveillance.
Supported by the Western Regional Center of Excellence for Biodefense and Emerging Infectious Diseases, NIH/NIAID U54 AI057156.