Abstract
BackgroundJapanese encephalitis virus (JEV) is the most important cause of epidemic encephalitis in most Asian regions. There is no specific treatment available for Japanese encephalitis, and vaccination is the only effective way to prevent JEV infection in humans and domestic animals. The purpose of this study is to establish a new mammalian cell line stably and efficiently expressing virus-like particle of JEV for potential use of JEV subunit vaccine.ResultsWe generated a new cell clone (BJ-ME cells) that stably produces a secreted form of Japanese encephalitis virus (JEV) virus-like particle (VLP). The BJ-ME cells were engineered by transfecting BHK-21 cells with a code-optimized cDNA encoding JEV prM and E protein expression plasmid. Cell line BJ-ME can stably produces a secreted form of Japanese encephalitis virus virus-like particle (JEV-VLP) which contains the JEV envelope glycoprotein (E) and membrane protein (M). The amount of JEV-VLP antigen released into the culture fluid of BJ-ME cells was as high as 15–20 μg/ml. JEV-VLP production was stable after multiple cell passages and 100% cell expression was maintained without detectable cell fusion or apoptosis. Cell culture fluid containing the JEV-VLP antigen could be harvested five to seven times continuously at intervals of 4–6 days while maintaining the culture. Mice immunized with the JEV-VLP antigen with or without adjuvant developed high titers of neutralizing antibodies and 100% protection against lethal JEV challenge.ConclusionThese results suggest that the recombinant JEV-VLP antigen produced by the BJ-ME cell line is an effective, safe and affordable subunit Japanese encephalitis vaccine candidate, especially for domestic animals such as pig and horse.
Highlights
Japanese encephalitis virus (JEV) is the most important cause of epidemic encephalitis in most Asian regions
When the fifth-passage BJ-ME cells were examined by indirect immunofluorescence and flow cytometry, over 94% of BJ-ME cells were positively stained with JEV E proteinspecific MAb 5E7
The results showed that virus-like particle (VLP) structure was observed in the endoplasmic reticulum of cells (Figure 3A,B), and this Japanese encephalitis virus virus-like particle (JEV-VLP) structure was further confirmed with purified JEV-VLP by negative staining electron microscopy observation (Figure 3C)
Summary
Japanese encephalitis virus (JEV) is the most important cause of epidemic encephalitis in most Asian regions. There is no specific treatment available for Japanese encephalitis, and vaccination is the only effective way to prevent JEV infection in humans and domestic animals. Japanese encephalitis virus (JEV) is the most important cause of epidemic encephalitis in most Asian regions, with about 35,000–50,000 cases of and 10,000 deaths from JEV infection reported annually [1]. A variety of animals are susceptible to JEV infection, but usually only humans, horses and pigs with infection exhibit symptoms [8,9,10,11]. Pigs are considered the most important amplification and reservoir hosts of JEV in endemic regions [7,13]. Infection of boars can cause acute testicular inflammation and hypospermia [14,15]
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