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Review
. 2011 Jan;21(1):18-31.
doi: 10.1002/rmv.678.

Efficient cell culture systems for hepatitis E virus strains in feces and circulating blood

Hiroaki Okamoto  1
Affiliations
Review

Efficient cell culture systems for hepatitis E virus strains in feces and circulating blood

Hiroaki Okamoto. Rev Med Virol. 2011 Jan.

Abstract

Attempts have been made to propagate hepatitis E virus (HEV) in primary hepatocyte culture and various other cultured cells. However, the replication ability of HEV recovered from culture media remains extremely low. Recently, efficient culture systems have been established in PLC/PRF/5 (hepatocellular carcinoma) and A549 (lung cancer) cell lines for HEV strains of genotypes 3 and 4 in our laboratory. They originated in fecal extracts from patients containing HEV RNA in extremely high-titers (10(7) copies/ml), and named the JE03-1760F (genotype 3) and HE-JF5/15F (genotype 4) strains, respectively. HEV RNA in culture supernatants reached 10(8) copies/ml in titer, and were transmitted successively through many passages. An infectious HEV cDNA clone (pJE03-1760F/wt) was constructed that has replication activity comparable to that of the wild-type JE03-1760F in feces. The ORF3 protein is indispensable for shedding HEV particles from cells in the reverse genetics system. HEV recovered from culture media, as well as circulating HEV, possess ORF3 proteins on the surface and are covered with cellular membranes, and therefore, ORF2 epitopes are buried in these particles. In contrast, HEV excreted into feces are naked nucleocapsids without a lipid layer or surface expression of the ORF3 protein. HEV in sera of patients with acute hepatitis E can infect and replicate in PLC/PRF/5 and A549 cells, with efficiency comparable to the circulating HEV RNA levels. High-efficiency cell culture systems for infectious viruses, thus developed, are expected to open up a new era and resolve many mysteries in the epidemiology, molecular biology, and treatment of HEV.

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