title: Rescue of a genotype 4 human hepatitis E virus from cloned cDNA and characterization of intergenotypic chimeric viruses in cultured human liver cells and in pigs.
abstract: Hepatitis E virus (HEV) is an important but extremely understudied human pathogen. Genotypes 1 and 2 are restricted to humans, whereas genotypes 3 and 4 are zoonotic, infecting both humans and pigs. This report describes, for the first time, the successful rescue of infectious HEV in vitro and in vivo from cloned cDNA of a genotype 4 human HEV (strain TW6196E). The complete genomic sequence of the TW6196E virus was determined and a full-length cDNA clone (pHEV-4TW) was assembled. Capped RNA transcripts from the pHEV-4TW clone were replication competent in Huh7 cells and infectious in HepG2/C3A cells. Pigs inoculated intrahepatically with capped RNA transcripts from pHEV-4TW developed an active infection, as evidenced by faecal virus shedding and seroconversion, indicating the successful rescue of infectious genotype 4 HEV and cross-species infection of pigs by a genotype 4 human HEV. To demonstrate the utility of the genotype 4 HEV infectious clone and to evaluate the potential viral determinant(s) for species tropism, four intergenotypic chimeric clones were constructed by swapping various genomic regions between genotypes 1 and 4, and genotypes 1 and 3. All four chimeric clones were replication competent in Huh7 cells, but only the two chimeras with sequences swapped between genotypes 1 and 4 human HEVs produced viruses capable of infecting HepG2/C3A cells. None of the four chimeras was able to establish a robust infection in pigs. The availability of a genotype 4 HEV infectious clone affords an opportunity to delineate the molecular mechanisms of HEV cross-species infection in the future.
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
authors: Laura
authors: L
authors: Córdoba
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
authors: Alicia R
authors: AR
authors: Feagins
authors: Department of Veterinary Diagnostic and Production Animal Medicine, Iowa State University, Ames, IA, USA.
authors: Tanja
authors: T
authors: Opriessnig
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
authors: Caitlin M
authors: CM
authors: Cossaboom
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
authors: Barbara A
authors: BA
authors: Dryman
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
authors: Yao-Wei
authors: YW
authors: Huang
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
authors: Xiang-Jin
authors: XJ
authors: Meng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D005243:Feces / Q000821:virology; D016679:Genome, Viral; D005838:Genotype; D056945:Hep G2 Cells; D016751:Hepatitis E / Q000821:virology; D016752:Hepatitis E virus / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000821:virology*; D008969:Molecular Sequence Data; D012315:RNA Caps / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics*; D017421:Sequence Analysis / Q000379:methods; D013552:Swine / Q000821:virology*; D014779:Virus Replication; D017201:Virus Shedding / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D012315:RNA Caps; D012367:RNA, Viral