Comprehensive mapping of a novel NS1 epitope conserved in flaviviruses within the Japanese encephalitis virus serocomplex

Hua, Rong-Hong; Liu, Li-Ke; Hong, Hao; Li, Yenan; Guo, Liping; Wang, Xiaolei; Qin, Cheng‐Feng; Bu, Zhigao

doi:10.1016/j.virusres.2014.03.001

Cited by 9 publications

(8 citation statements)

References 23 publications

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“…It is a characteristic feature of antibodies against the E and NS1 proteins of JEV to cross-react with other viruses in the JE serogroup as well as with some viruses outside the JE serogroup, such as the DENV [1,4]. The anti-flavivirus group-antigen antibody (4G2) cross reactivity is confirmed in this study since 4G2 antibody detected the E proteins of JEV, WNV and four serotypes of DENV.…”

Section: Discussionsupporting

confidence: 71%

“…These cross-reactive responses make the interpretation of serological tests such as, enzyme-linked immunosorbent assay (ELISA) and neutralization tests, difficult [3]. Antibodies against the E protein of JEV could cross-react with members of JEV serogroup as well as with some viruses outside the JEV serogroup, such as DENV [4]. NS1 was found to be more specific than the E protein for serological testing and some antibody-based epitope-blocking ELISAs have been developed to differentiate between JEV, DENV and WNV infections in humans [5].…”

Section: Introductionmentioning

confidence: 99%

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Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

Kaufusi

Tseng

Kelley

et al. 2020

Viruses

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Studies investigating West Nile virus (WNV) NS4B protein function are hindered by the lack of an antibody recognizing WNV NS4B protein. Few laboratories have produced WNV NS4B antibodies, and none have been shown to work consistently. In this report, we describe a NS4B antibody against Japanese encephalitis virus (JEV) NS4B protein that cross-reacts with the NS4B protein of WNV but not of dengue virus (DENV). This JEV NS4B antibody not only recognizes WNV NS4B in infected cells, but also recognizes the NS4B protein expressed using transfection. It is evident from this data that the JEV NS4B antibody is specific to NS4B of WNV but not to NS4B of the four DENV serotypes. The specificity of this antibody may be due to the notable differences that exist between the amino acid sequence identity and antigenic relationships within the NS4B protein of the WNV, DENV, and JEV.

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Section: Discussionsupporting

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

Kaufusi

Tseng

Kelley

et al. 2020

Viruses

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“…The purified recombinant protein MBP-E2 was used as an immunogen in mice. Hybridomas secreting anti-E2 antibodies were generated according to the procedure described previously [20]–[22]. Ascitic fluid was generated in pristane-primed BALB/c mice.…”

Section: Methodsmentioning

confidence: 99%

“…For N -linked glycosylation analysis, the concentrated sample was digested with peptide: N -glycosidase F (PNGase, New England Biolabs) according to the manufacturer's instruction. The samples were analyzed by Western blot as described previously [22]. Briefly, samples were separated on Any kD Resolving Gel (Mino-PROTEAN TGX Precast Gels; Bio-Rad Laboratories, CA).…”

Section: Methodsmentioning

confidence: 99%

Generation and Efficacy Evaluation of Recombinant Classical Swine Fever Virus E2 Glycoprotein Expressed in Stable Transgenic Mammalian Cell Line

Hua

Hong

et al. 2014

PLoS ONE

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Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), which is a highly contagious swine disease that causes significant economic loses to the pig industry worldwide. The envelope E2 glycoprotein of CSFV is the most important viral antigen in inducing protective immune response against CSF. In this study, we generated a mammalian cell clone (BCSFV-E2) that could stably produce a secreted form of CSFV E2 protein (mE2). The mE2 protein was shown to be N-linked glycosylated and formed a homodimer. The vaccine efficacy of mE2 was evaluated by immunizing pigs. Twenty-five 6-week-old Landrace piglets were randomly divided into five groups. Four groups were intramuscularly immunized with mE2 emulsified in different adjuvants twice at four-week intervals. One group was used as the control group. All mE2-vaccinated pigs developed CSFV-neutralizing antibodies two weeks after the first vaccination with neutralizing antibody titers ranging from 1∶40 to 1∶320. Two weeks after the booster vaccination, the neutralizing antibody titers increased greatly and ranged from 1∶10,240 to 1∶81,920. At 28 weeks after the booster vaccine was administered, the neutralizing antibody titers ranged from 1∶80 to 1∶10240. At 32 weeks after the first vaccination, pigs in all the groups were challenged with a virulent CSFV strain at a dose of 1×105 TCID50. At two weeks after the challenge, all the mE2-immunized pigs survived and exhibited no obvious symptoms of CSF. The neutralizing antibody titer at this time was 20,480. Unvaccinated pigs in the control group exhibited symptoms of CSF 3–4 days after challenge and were euthanized from 7–9 days after challenge when the pigs became moribund. These results indicate that the mE2 is a good candidate for the development of a safe and effective CSFV subunit vaccine.

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“…5E7 and 5B10 which were generated with purified BJ-ME antigen according to the procedure described previously [44-46]. Briefly MAb 5B10 (2 μg/ml) was coated on 96-well plates overnight at 4°C, and blocked with 5% skimmed dried milk for 2 h at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Generation and characterization of a new mammalian cell line continuously expressing virus-like particles of Japanese encephalitis virus for a subunit vaccine candidate

Hua

Chen

et al. 2014

BMC Biotechnol

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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.

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Comprehensive mapping of a novel NS1 epitope conserved in flaviviruses within the Japanese encephalitis virus serocomplex

Cited by 9 publications

References 23 publications

Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

Generation and Efficacy Evaluation of Recombinant Classical Swine Fever Virus E2 Glycoprotein Expressed in Stable Transgenic Mammalian Cell Line

Generation and characterization of a new mammalian cell line continuously expressing virus-like particles of Japanese encephalitis virus for a subunit vaccine candidate

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