A Recombinant Chimeric Cedar Virus-Based Surrogate Neutralization Assay Platform for Pathogenic Henipaviruses
Moushimi Amaya,
Randy Yin,
Lianying Yan,
Viktoriya Borisevich,
Bishwo N. Adhikari,
Andrew Bennett,
Francisco Malagon,
Regina Z. Cer,
Kimberly A. Bishop-Lilly,
Antony S. Dimitrov,
Robert W. Cross,
Thomas W. Geisbert,
Christopher C. Broder
Affiliations
Moushimi Amaya
Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814, USA
Randy Yin
Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814, USA
Lianying Yan
Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814, USA
Viktoriya Borisevich
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA
Bishwo N. Adhikari
Genomics and Bioinformatics Department, Biological Defense Research Directorate, Naval Medical Research Command–Frederick, Fort Detrick, Frederick, MD 21702, USA
Andrew Bennett
Defense Threat Reduction Agency, Fort Belvoir, VA 22060, USA
Francisco Malagon
Defense Threat Reduction Agency, Fort Belvoir, VA 22060, USA
Regina Z. Cer
Genomics and Bioinformatics Department, Biological Defense Research Directorate, Naval Medical Research Command–Frederick, Fort Detrick, Frederick, MD 21702, USA
Kimberly A. Bishop-Lilly
Genomics and Bioinformatics Department, Biological Defense Research Directorate, Naval Medical Research Command–Frederick, Fort Detrick, Frederick, MD 21702, USA
Antony S. Dimitrov
Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814, USA
Robert W. Cross
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA
Thomas W. Geisbert
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA
Christopher C. Broder
Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814, USA
The henipaviruses, Nipah virus (NiV), and Hendra virus (HeV) can cause fatal diseases in humans and animals, whereas Cedar virus is a nonpathogenic henipavirus. Here, using a recombinant Cedar virus (rCedV) reverse genetics platform, the fusion (F) and attachment (G) glycoprotein genes of rCedV were replaced with those of NiV-Bangladesh (NiV-B) or HeV, generating replication-competent chimeric viruses (rCedV-NiV-B and rCedV-HeV), both with and without green fluorescent protein (GFP) or luciferase protein genes. The rCedV chimeras induced a Type I interferon response and utilized only ephrin-B2 and ephrin-B3 as entry receptors compared to rCedV. The neutralizing potencies of well-characterized cross-reactive NiV/HeV F and G specific monoclonal antibodies against rCedV-NiV-B-GFP and rCedV-HeV-GFP highly correlated with measurements obtained using authentic NiV-B and HeV when tested in parallel by plaque reduction neutralization tests (PRNT). A rapid, high-throughput, and quantitative fluorescence reduction neutralization test (FRNT) using the GFP-encoding chimeras was established, and monoclonal antibody neutralization data derived by FRNT highly correlated with data derived by PRNT. The FRNT assay could also measure serum neutralization titers from henipavirus G glycoprotein immunized animals. These rCedV chimeras are an authentic henipavirus-based surrogate neutralization assay that is rapid, cost-effective, and can be utilized outside high containment.
