Arenaviruses include several causative agents of Hemorrhagic Fever (HF) disease in humans that are associated with high morbidity and significant mortality. Moreover, weaponized forms of arenaviruses pose a serious threat as agents of bioterrorism. Public health concerns posed by arenaviruses are aggravated by the lack of licensed vaccines and by current anti-arenavirus therapy limited to the off-label use of ribavirin, which is only partially effective and often associated with severe side effects. Limitations in the study of HF arena-viruses include the manipulation of live forms of these agents under BioSafety Level (BSL) 4 laboratories and the requirement of secondary assays for detection of the virus. Development of valid single-cycle infectious surrogates that allow the study of HF arenavirus under less-strict BSL2 facilities and that express a reporter gene for easy viral detection will facilitate the identification of prophylactic and therapeutic strategies using safe, sensitive and specific screening assays compatible with High Throughput Screening (HTS) technologies. Furthermore, they will also represent promising vaccine approaches that circumvent concerns about potential out-of-control replication of attenuated vaccine strains that might lead to disease. Recently, we have described, for the first time, the generation of a single-cycle infectious, reporter-expressing, Old World Lymphocytic Choriomeningitis Virus (LCMV) where we replaced the viral glycoprotein (GP) with a reporter green fluorescent protein (GFP), rLCMV?GP/GFP. Infectious virus was achieved via genetic trans- complementation with cell lines constitutively expressing LCMV GP. This system allowed us to study multiple aspects of the virus and to develop screening assays for detection and quantification of neutralizing antibodies and identification of viral inhibitors. We were also able to extend our work to Lassa virus (LASV), an HF member of the Old World arenavirus, by generating LASV GP-expressing cell lines and LASV GP-pseudotyped rLCMV?GP/GFP. Since Old World and New World arena-viruses differ in their reservoirs, cellular receptor use for viral entry, and RNA genome composition, the development of valid surrogates to study New World arenavirus is imperative to provide researchers a way to study all aspects of the virus biology, to identify antivirals, and to develop vaccines against New World arenaviruses under less strict biosafety laboratories. In this application we propose to expand our recently described technology to the study of New World arenavirus by generating a single-cycle infectious, reporter-expressing, attenuated vaccine strain Candid#1 (rCan?GP/GFP).

Public Health Relevance

Arenaviruses are important viral pathogens that cause common and in some case severe infections in humans. Among them, Hemorrhagic Fever (HF) arenaviruses represent a serious public health problem as well as a threat as agents of bioterrorism. No licensed anti-arenavirus vaccines are available, and current anti-arenavirus therapy is limited to the use of ribavirin, which is only partially effective and often associated with severe side effects. Our goal is to generate a valid surrogate system that facilitates the study of HF New World members in the family under less-strict biosafety conditions by generating a single-cycle infectious, reporter-expressing, attenuated vaccine strain Candid#1.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Small Research Grants (R03)
Project #
5R03AI099681-02
Application #
8484790
Study Section
Virology - A Study Section (VIRA)
Program Officer
Repik, Patricia M
Project Start
2012-07-01
Project End
2014-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
2
Fiscal Year
2013
Total Cost
$76,750
Indirect Cost
$26,750
Name
University of Rochester
Department
Microbiology/Immun/Virology
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
Martínez-Sobrido, Luis; Cheng, Benson Yee Hin; de la Torre, Juan Carlos (2016) Reverse Genetics Approaches to Control Arenavirus. Methods Mol Biol 1403:313-51
Yang, Hongmei; Baker, Steven F; González, Mario E et al. (2016) An improved method for estimating antibody titers in microneutralization assay using green fluorescent protein. J Biopharm Stat 26:409-20
Martinez-Sobrido, Luis; de la Torre, Juan Carlos (2016) Novel strategies for development of hemorrhagic fever arenavirus live-attenuated vaccines. Expert Rev Vaccines 15:1113-21
Cheng, Benson Yee Hin; Ortiz-Riaño, Emilio; de la Torre, Juan Carlos et al. (2015) Arenavirus Genome Rearrangement for the Development of Live Attenuated Vaccines. J Virol 89:7373-84
Baker, Steven F; Nogales, Aitor; Santiago, Felix W et al. (2015) Competitive detection of influenza neutralizing antibodies using a novel bivalent fluorescence-based microneutralization assay (BiFMA). Vaccine 33:3562-70
Pythoud, Christelle; Rothenberger, Sylvia; Martínez-Sobrido, Luis et al. (2015) Lymphocytic Choriomeningitis Virus Differentially Affects the Virus-Induced Type I Interferon Response and Mitochondrial Apoptosis Mediated by RIG-I/MAVS. J Virol 89:6240-50
Cheng, Benson Yee Hin; Ortiz-Riaño, Emilio; Nogales, Aitor et al. (2015) Development of live-attenuated arenavirus vaccines based on codon deoptimization. J Virol 89:3523-33
DeVito, Stefanie Renee; Ortiz-Riaño, Emilio; Martínez-Sobrido, Luis et al. (2014) Cytomegalovirus-mediated activation of pyrimidine biosynthesis drives UDP-sugar synthesis to support viral protein glycosylation. Proc Natl Acad Sci U S A 111:18019-24
Ramon, Sesquile; Baker, Steven F; Sahler, Julie M et al. (2014) The specialized proresolving mediator 17-HDHA enhances the antibody-mediated immune response against influenza virus: a new class of adjuvant? J Immunol 193:6031-40
Guo, Hailong; Baker, Steven F; Martínez-Sobrido, Luis et al. (2014) Induction of CD8 T cell heterologous protection by a single dose of single-cycle infectious influenza virus. J Virol 88:12006-16

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