Generation and characterization of genetically and antigenically diverse infectious clones of dengue virus serotypes 1–4. Issue 1 (31st December 2022)

• Record Type: Journal Article
• Authors: Tamura, Tomokazu; Zhang, Jiayu; Madan, Vrinda; Biswas, Abhishek; Schwoerer, Michael P.; Cafiero, Thomas R.; Heller, Brigitte L.; Wang, Wei; Ploss, Alexander
• Is Part Of: Emerging microbes & infections. Volume 11:Issue 1(2022)
• Page Start: 227
• Page End: 239
• Publication Date: 2022-12-31
• Subjects: Dengue -- dengue virus -- host tropism -- animal models -- host response; Medical microbiology -- Periodicals; Communicable diseases -- Periodicals; Infection -- Periodicals; 616.9041
• DOI: 10.1080/22221751.2021.2021808
• Languages: English
• ISSNs: 2222-1751

Abstract

ABSTRACT: Dengue is caused by four genetically distinct viral serotypes, dengue virus (DENV) 1-4. Following transmission by Aedes mosquitoes, DENV can cause a broad spectrum of clinically apparent disease ranging from febrile illness to dengue hemorrhagic fever and dengue shock syndrome. Progress in the understanding of different dengue serotypes and their impacts on specific host-virus interactions has been hampered by the scarcity of tools that adequately reflect their antigenic and genetic diversity. To bridge this gap, we created and characterized infectious clones of DENV1–4 originating from South America, Africa, and Southeast Asia. Analysis of whole viral genome sequences of five DENV isolates from each of the four serotypes confirmed their broad genetic and antigenic diversity. Using a modified circular polymerase extension reaction (CPER), we generated de novo viruses from these isolates. The resultant clones replicated robustly in human and insect cells at levels similar to those of the parental strains. To investigate in vivo properties of these genetically diverse isolates, representative viruses from each DENV serotype were administered to NOD Rag1 −/−, IL2rg null Flk2 −/− (NRGF) mice, engrafted with components of a human immune system. All DENV strains tested resulted in viremia in humanized mice and induced cellular and IgM immune responses. Collectively, we describe here a workflow for rapidly generating de novo infectious clones of DENV – and conceivably other RNA viruses. The infectious clones described here are a valuable resource for reverse genetic studies and for characterizing host responses to DENV in vitro and in vivo .