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Generation and Characterization of a Replication-Competent Human Adenovirus Type 55 Encoding EGFP
Wei Li1, Yuehong Chen1, Ye Feng1
1State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.
Viruses
|May 27, 2023
Summary
A new infectious Human adenovirus 55 (HAdV-55) virus, rAdv55-dE3-EGFP, was created for rapid antiviral screening. This tool aids in quantifying neutralizing antibodies and developing new treatments for acute respiratory disease (ARD).
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Human adenovirus 55 (HAdV-55) outbreaks cause significant acute respiratory disease (ARD) in civilian and military populations.
- Developing antiviral inhibitors and quantifying neutralizing antibodies necessitates rapid viral infection monitoring systems.
- Infectious virus production from plasmids is crucial for creating experimental monitoring systems.
Purpose of the Study:
- To construct a full-length infectious cDNA clone of HAdV-55 for generating a recombinant virus.
- To develop a rapid, high-throughput assay for HAdV-55 neutralization testing and antiviral screening.
Main Methods:
- Bacteria-mediated recombination was used to create the pAd55-FL clone of the HAdV-55 genome.
- A green fluorescent protein (GFP) expression cassette replaced the E3 region, creating pAd55-dE3-EGFP.
- The recombinant virus rAdv55-dE3-EGFP was rescued and characterized for stability and replication.
Main Results:
- The recombinant virus rAdv55-dE3-EGFP is genetically stable and replicates similarly to wild-type HAdV-55 in cell culture.
- rAdv55-dE3-EGFP effectively quantifies neutralizing antibodies in sera, aligning with traditional microneutralization assays.
- The assay demonstrated utility in antiviral screening using A549 cells.
Conclusions:
- The rAdv55-dE3-EGFP virus is a reliable tool for rapid HAdV-55 neutralization testing.
- This high-throughput assay facilitates efficient antiviral screening against HAdV-55.
- The developed system supports the development of countermeasures against HAdV-55-induced ARD.
