Establishment of hepatitis E virus infection‐permissive and ‐non‐permissive human hepatoma PLC/PRF/5 subclones

Abstract

PLC/PRF/5 cells show limited permissiveness, meaning that almost all subclones are permissive; however, some subclones do not exhibit permissiveness for hepatitis E virus (HEV) infection. In this study, the single-cell cloning of PLC/PRF/5 was performed and heterogeneous subclones characterized. Notably, the efficiency of intracellular virus replication did not correlate with the permissiveness for HEV infection. However, as well as binding permissive subclones, virus-like particles bound non-permissive subclones on various levels, suggesting that these subclones have some deficiencies in the attachment and entry steps of infection. Our data would be useful for investigating the HEV life cycle.

Robust hepatitis E virus infection and transcriptional response in human hepatocytes

“…S4A). To universalize our results, we analyzed another gt 3 strain (HEV83-2-27) that lacks the insertion in the hypervariable region (HVR) (24). Introduction of the mutation in a Gaussia reporter replicon revealed no enhancement of viral fitness as previously demonstrated for the p6 strain (13,22), neither when assessing HEV RNA copy numbers (SI Appendix, Fig.…”

Enhanced Replication of Hepatitis E Virus Strain 47832c in an A549-Derived Subclonal Cell Line

“…In this context, it should also be considered that the A549 cell lines used in different laboratories may be heterogeneous and therefore show different efficiencies of HEV replication. Subclonal cell lines with differing efficiency of HEV replication have also been described very recently for the PLC/PRF/5 cell line [21], thus indicating that this phenomenon is not unique for A549 cells. The selected subclonal cell lines may later be used in improved HEV cell culture systems.…”

Hepatitis E Virus Lifecycle and Identification of 3 Forms of the ORF2 Capsid Protein

“…Infection of humanized mice. Primary human hepatocytes were transplanted into homozygous 1 uPA +/+ -SCID mice, as previously described 8 . Humanized mice were inoculated via intrasplenic route 2 with HEV-p6 purified on iodixanol gradient (fraction6, F6p6, 8.7x10 7 IU/mouse) or with the same 3 volume of fraction6 from a control gradient prepared with concentrated supernatant of non-transfected 4 PLC3 cells (F6 control).…”

“…Peptides were filtered out according to the cutoff ion score > 25 and a minimal size of 7 amino acid residues. 5 1.0x10 5 20.2 ORF2g/ORF2c P5 3f 7.0x10 5 3.5x10 5 ND -P6 3c 1.6x10 8 1.6x10 7 19.4 ORF2g/ORF2c P7 3c 1.2x10 7 1.2x10 6 18.6 ORF2g/ORF2c P8 3f 1.2x10 5 0.6x10 5 5 -P9 3f 1.5x10 5 0.7x10 5 19.7 ORF2g/ORF2c P10 3f 1.2x10 6 0.6x10 6 18.9 ORF2g/ORF2c a in HEV RNA copies/ml b in HEV RNA copies c in S/CO values. Samples were diluted 100 times in PBS and quantified with the Wantaï HEV-Ag-ELISA kit.…”