Development of Live-Attenuated Virus Vaccine Platform Against Hemorrhagic Fever Causing Arenaviruses
Abstract
Topic Area: The proposed project relates to the Topic Area “Vaccine Development for Infectious Disease”. The proposal seeks to develop a live-attenuated vaccine (LAV) platform to combat infections by the main hemorrhagic fever (HF)-causing mammarenaviruses and to provide the basis for a universal mammarenavirus vaccine platform to combat diseases caused by these important human pathogens. The Old World (OW) mammarenavirus Lassa (LASV) in West Africa and the New World (NW) mammarenavirus Junin (JUNV) in Argentina cause hemorrhagic fever (HF) diseases in humans that are associated with high morbidity and mortality. Notably, increased traveling has led to the importation of arenaviral HF disease cases into non-endemic metropolitan areas around the globe. Moreover, HF-causing mammarenavirus, including LASV and JUNV, pose a credible biodefense threat and are classified as NIAID [National Institute of Allergy and Infectious Diseases] Category A priority pathogens. Concerns posed by HF-causing mammarenaviruses are aggravated by the lack of Food and Drug Administration (FDA)-licensed vaccines and current anti-arenaviral therapy being limited to the off-label use of ribavirin that is only partially effective and associated with side effects. The significance of HF-causing LASV and JUNV in human health and biodefense readiness, together with the limited existing armamentarium to combat them, highlight the importance of developing effective countermeasures to combat HF-causing mammarenavirus infections in humans. The central goal of this application is to demonstrate that recombinant forms of the prototypic mammarenavirus lymphocytic choriomeningitis virus (LCMV) containing a codon deoptimized (CD) nucleoprotein (NP) expressing glycoproteins (GP) of HF-causing LASV and JUNV (rLCMV/NPCD/GPHF) can be used for the development of a safe and protective individual or blended live-attenuated vaccine (LAV) to combat HF disease caused by LASV and JUNV infections. We have obtained preliminary data that support the concept and experimental approach of our proposal as well as the feasibility of implementing our approach for the successful development of LAV for the treatment of HF-causing LASV and JUNV. (1) We have shown that mice immunized with a rLCMV/NPCD remain free of clinical symptoms and cleared the virus, but were protected against a subsequent lethal challenge with rLCMV/WT. (2) In FDA-approved for production of therapeutics BHK-21 cells, rLCMV/NPCD grows to titers compatible with vaccine manufacturing and production. (3) rLCMV/NPCD contains a large number (197) of silent mutations that pose an insurmountable barrier for reversion to a virulent strain. (4) We have obtained preliminary data that demonstrate the genetic and phenotypic stability of our rLCMV/NPCD. (5) We have successfully rescued rLCMVs where the viral GP was substituted by that of LASV or JUNV and shown they grow to high titers in cultured cells. (6) We have generated LASV and JUNV GP-expressing stable cell lines that are able to complement, to levels comparable to LCMV GP, a recombinant LCMV where the viral GP has been substituted by GFP (rLCMVDGP/GFP). (7) We have developed robust reverse genetics approaches that allow us to rescue recombinant wild-type (WT) as well as mutant forms of LCMV. (8) Likewise, we are very familiar with the experimental procedures to characterize rLCMVs generated by reverse genetics in vitro and in vivo, including the model of LCMV induced fatal lymphocytic choriomeningitis (LCM) disease in mice. (9) We are very familiar with measuring adaptive T cell and total and neutralizing antibody responses in mice immunized with rLCMV and the assays to evaluate T- and B-cell activation are well established in our laboratories and we have extensive experience with them. (10) Finally, Dr. Carrion at Texas Biomedical Research Institute, is highly experience in the use of guinea pigs as models for HF-causing mammarenaviruses, including LA
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Nov 19, 2019
- Source ID
- W81XWH1910496
Entities
People
- Luis Martinez-Sobrido
Organizations
- United States Army
- University of Rochester