title: Three amino acid mutations (F51L, T59A, and S390L) in the capsid protein of the hepatitis E virus collectively contribute to virus attenuation.
abstract: Hepatitis E virus (HEV) is an important but extremely understudied human pathogen, and the mechanisms of HEV replication and pathogenesis are largely unknown. We previously identified an attenuated genotype 3 HEV mutant (pSHEV-1) containing three unique amino acid mutations (F51L, T59A, and S390L) in the capsid protein. To determine the role of each of these mutations, we constructed three HEV single mutants (rF51L, rT59A, and rS390L) which were all found to be replication competent in Huh7 liver cells. To determine the pathogenicities of the mutants, we utilized the specific-pathogen-free (SPF) pig model for HEV and a unique inoculation procedure that bypasses the need for propagating infectious HEV in vitro. A total of 60 pigs were intrahepatically inoculated, via an ultrasound-guided technique, with in vitro-transcribed full-length capped RNA transcripts from the infectious clones of each single mutant, the pSHEV-1 triple mutant, wild-type pSHEV-3, or phosphate-buffered saline (PBS) buffer (n = 10). The results showed that the F51L mutation partially contributed to virus attenuation, whereas the T59A and S390L mutations resulted in more drastic attenuation of HEV in pigs, as evidenced by a significantly lower incidence of viremia, a delayed appearance and shorter duration of fecal virus shedding and viremia, and lower viral loads in liver, bile, and intestinal content collected at three different necropsy times. The results indicate that the three mutations in the capsid protein collectively contribute to HEV attenuation. This study has important implications for developing a modified live-attenuated vaccine against HEV.
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA.
authors: Laura
authors: L
authors: Córdoba
authors: Yao-Wei
authors: YW
authors: Huang
authors: Tanja
authors: T
authors: Opriessnig
authors: Kylie K
authors: KK
authors: Harral
authors: Nathan M
authors: NM
authors: Beach
authors: Carla V
authors: CV
authors: Finkielstein
authors: Suzanne U
authors: SU
authors: Emerson
authors: Xiang-Jin
authors: XJ
authors: Meng
mesh_terms: D000596:Amino Acids / Q000235:genetics*; D000818:Animals; D001769:Blood / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004195:Disease Models, Animal; D005243:Feces / Q000821:virology; D016751:Hepatitis E / Q000473:pathology / Q000821:virology; D016752:Hepatitis E virus / Q000235:genetics / Q000472:pathogenicity*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D020125:Mutation, Missense; D013552:Swine; D013553:Swine Diseases / Q000473:pathology / Q000821:virology; D014766:Viremia; D014774:Virulence; D017201:Virus Shedding
chemical_list: D000596:Amino Acids; D036022:Capsid Proteins