Enhanced Replication of Hepatitis E Virus Strain 47832c in an A549-Derived Subclonal Cell Line.
Hepatitis E virus (HEV) is a human pathogen with increasing importance. The lack of efficient cell culture systems hampers systematic studies on its replication cycle, virus neutralization and inactivation. Here, several cell lines were inoculated with the HEV genotype 3c strain 47832c, previously isolated from a chronically infected transplant patient. At 14 days after inoculation the highest HEV genome copy numbers were found in A549 cells, followed by PLC/PRF/5 cells, whereas HepG2/C3A, Huh-7 Lunet BLR and MRC-5 cells only weakly supported virus replication. Inoculation of A549-derived subclone cell lines resulted in most cases in reduced HEV replication. However, the subclone A549/D3 was susceptible to lower virus concentrations and resulted in higher virus yields as compared to parental A549 cells. Transcriptome analysis indicated a downregulation of genes forcarcinoembryonic antigen-related cell adhesion molecules(CEACAM)5and6, and an upregulation of thesyndecan 2(SDC2) gene in A549/D3 cells compared to A549 cells. However, treatment of A549/D3 cells or A549 cells with CEACAM- or syndecan 2-specific antisera did not influence HEV replication. The results show that cells supporting more efficient HEV replication can be selected from the A549 cell line. The specific mechanisms responsible for the enhanced replication remain unknown.
Institute of Clinical Microbiology and Hygiene, 93053 Regensburg, Germany. Mathias.Schemmerer@klinik.uni-regensburg.de.
Mathias
M
Schemmerer
German Federal Institute for Risk Assessment, Department of Biological Safety, 12277 Berlin, Germany. Silke.Apelt@bfr.bund.de.
Silke
S
Apelt
German Federal Institute for Risk Assessment, Department of Biological Safety, 12277 Berlin, Germany. Eva.Trojnar@bfr.bund.de.
Eva
E
Trojnar
Friedrich-Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, 17493 Greifswald-Insel Riems, Germany. Rainer.Ulrich@fli.bund.de.
Rainer G
RG
Ulrich
Institute of Clinical Microbiology and Hygiene, 93053 Regensburg, Germany. Juergen.Wenzel@klinik.uni-regensburg.de.
Jürgen J
JJ
Wenzel
German Federal Institute for Risk Assessment, Department of Biological Safety, 12277 Berlin, Germany. Reimar.Johne@bfr.bund.de.
Reimar
R
Johne
A549; CEACAM; cell culture; hepatitis E virus; syndecan
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