Display of the antigenic region of Nipah virus nucleocapsid protein on hepatitis B virus capsid

Yap, Wei Boon; Tey, Beng Ti; Alitheen, Noorjahan Banu; Tan, Wen Siang

doi:10.1016/j.jbiosc.2011.09.007

Cited by 9 publications

(9 citation statements)

References 11 publications

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“…VLPs formed by the capsid proteins of viruses such as HBV (25,27,32) and human papillomavirus (33) and bacteriophages (8,34) have been widely used for displaying foreign epitopes for vaccine development. The carriers enhance the antigenicity of the fused epitopes (35)(36)(37), which are often found to be inefficient in eliciting immune responses.…”

Section: Discussionmentioning

confidence: 99%

“…The recombinant MrNV capsid protein expressed in Escherichia coli self-assembled into VLPs in the absence of other viral proteins (18,19). VLPs have been widely used for various purposes, including drug delivery (20,21), gene therapy (22), vaccine development (23)(24)(25)(26)(27), and display of epitopes (28). Therefore, it is hypothesized that the VLPs of MrNV capsid protein can be used to display foreign epitopes.…”

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confidence: 99%

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Induction of Humoral and Cell-Mediated Immune Responses by Hepatitis B Virus Epitope Displayed on the Virus-Like Particles of Prawn Nodavirus

Yong

Yeap

Goh

et al. 2015

Appl Environ Microbiol

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Hepatitis B virus (HBV) is a deadly pathogen that has killed countless people worldwide. Saccharomyces cerevisiae-derived HBV vaccines based upon hepatitis B surface antigen (HBsAg) is highly effective. However, the emergence of vaccine escape mutants due to mutations on the HBsAg and polymerase genes has produced a continuous need for the development of new HBV vaccines. In this study, the "a" determinant within HBsAg was displayed on the recombinant capsid protein of Macrobrachium rosenbergii nodavirus (MrNV), which can be purified easily in a single step through immobilized metal affinity chromatography (IMAC). The purified protein self-assembled into virus-like particles (VLPs) when observed under a transmission electron microscope (TEM). Immunization of BALB/c mice with this chimeric protein induced specific antibodies against the "a" determinant. In addition, it induced significantly more natural killer and cytotoxic T cells, as well as an increase in interferon gamma (IFN-␥) secretion, which are vital for virus clearance. Collectively, these findings demonstrated that the MrNV capsid protein is a potential carrier for the HBV "a" determinant, which can be further extended to display other foreign epitopes. This paper is the first to report the application of MrNV VLPs as a novel platform to display foreign epitopes. Hepatitis B virus (HBV) is a partially double-stranded DNA virus that belongs to the family Hepadnaviridae. It is known to be the major cause of liver-associated diseases, including liver cirrhosis and hepatocellular carcinoma. Approximately 2 billion people worldwide have been infected by HBV, among which about 350 million chronically infected people serve as a reservoir for the virus (1, 2). To date, there is no effective treatment for HBV infection despite intensive studies on antiviral drugs. Thus, preventive measures, including immunization with HBV vaccine, remain necessary.HBV surface antigens (HBsAg) purified from the sera of infected patients have been used for immunization against HBV infection since 1981. The full-length HBsAg gene has three different start codons and a common stop codon and encodes HBsAg of different lengths (3). The longest is the large HBsAg (L-HBsAg), followed by the middle HBsAg (M-HBsAg), and the smallest is the small HBsAg (S-HBsAg). L-HBsAg is composed of 389 or 400 amino acids (aa), depending on the virus genotypes. It contains the pre-S1 region (108 or 119 aa), followed by the pre-S2 region (55 aa) and S-HBsAg (226 aa). The N-terminal end of M-HBsAg harbors the pre-S2 region at its S-HBsAg. The S-HBsAg contains 226 aa in the absence of the pre-S1 and pre-S2 regions.The HBsAg in serum self-assemble into noninfectious 22-nm spherical or filamentous particles, along with lipid components (4), which are capable of inducing neutralizing antibodies against HBV infection. Although highly effective, the use of plasma-derived HBsAg for vaccination is limited by the production cost, as well as the possibility of contamination by other infectious pathogens present in...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Induction of Humoral and Cell-Mediated Immune Responses by Hepatitis B Virus Epitope Displayed on the Virus-Like Particles of Prawn Nodavirus

Yong

Yeap

Goh

et al. 2015

Appl Environ Microbiol

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“…VLPs have been widely used for displaying foreign epitopes, for instance the VLPs of human papilloma virus ( Matic et al, 2011 ), HBV ( Ibañez et al, 2013 ; Murray & Shiau, 1999 ; Yap et al, 2012 ), as well as bacteriophages ( Hashemi et al, 2012 ; Kok et al, 2002 ; Tan et al, 2005 ; Wan et al, 2001 ). VLPs are known to enhance the immunogenicity of small epitopes displayed on the particles ( Murata et al, 2003 ; Quan et al, 2008 ).…”

Section: Survey Methodologymentioning

confidence: 99%

Advances in the study of nodavirus

Yong

Yeap

Omar

et al. 2017

PeerJ

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Nodaviruses are small bipartite RNA viruses which belong to the family of Nodaviridae. They are categorized into alpha-nodavirus, which infects insects, and beta-nodavirus, which infects fishes. Another distinct group of nodavirus infects shrimps and prawns, which has been proposed to be categorized as gamma-nodavirus. Our current review focuses mainly on recent studies performed on nodaviruses. Nodavirus can be transmitted vertically and horizontally. Recent outbreaks have been reported in China, Indonesia, Singapore and India, affecting the aquaculture industry. It also decreased mullet stock in the Caspian Sea. Histopathology and transmission electron microscopy (TEM) are used to examine the presence of nodaviruses in infected fishes and prawns. For classification, virus isolation followed by nucleotide sequencing are required. In contrast to partial sequence identification, profiling the whole transcriptome using next generation sequencing (NGS) offers a more comprehensive comparison and characterization of the virus. For rapid diagnosis of nodavirus, assays targeting the viral RNA based on reverse-transcription PCR (RT-PCR) such as microfluidic chips, reverse-transcription loop-mediated isothermal amplification (RT-LAMP) and RT-LAMP coupled with lateral flow dipstick (RT-LAMP-LFD) have been developed. Besides viral RNA detections, diagnosis based on immunological assays such as enzyme-linked immunosorbent assay (ELISA), immunodot and Western blotting have also been reported. In addition, immune responses of fish and prawn are also discussed. Overall, in fish, innate immunity, cellular type I interferon immunity and humoral immunity cooperatively prevent nodavirus infections, whereas prawns and shrimps adopt different immune mechanisms against nodavirus infections, through upregulation of superoxide anion, prophenoloxidase, superoxide dismutase (SOD), crustin, peroxinectin, anti-lipopolysaccharides and heat shock proteins (HSP). Potential vaccines for fishes and prawns based on inactivated viruses, recombinant proteins or DNA, either delivered through injection, oral feeding or immersion, are also discussed in detail. Lastly, a comprehensive review on nodavirus virus-like particles (VLPs) is presented. In recent years, studies on prawn nodavirus are mainly focused on Macrobrachium rosenbergii nodavirus (MrNV). Recombinant MrNV VLPs have been produced in prokaryotic and eukaryotic expression systems. Their roles as a nucleic acid delivery vehicle, a platform for vaccine development, a molecular tool for mechanism study and in solving the structures of MrNV are intensively discussed.

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“…The self‐assembly capability of HBcAg can be manipulated to display foreign epitopes on the VLP. These epitopes derived from foot‐and‐mouth disease virus, plasmodium spp ., influenza virus, and anthrax protective antigen . Due to its potential applications in the development of multicomponent vaccines and diagnostic reagents, hence it is of importance to establish an effective method to purify and to separate the HBcAg large and small particles.…”

Section: Introductionmentioning

confidence: 99%

Native agarose gel electrophoresis and electroelution: A fast and cost‐effective method to separate the small and large hepatitis B capsids

et al. 2012

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Hepatitis B core antigen (HBcAg) expressed in Escherichia coli is able to self-assemble into large and small capsids comprising 240 (triangulation number T = 4) and 180 (triangulation number T = 3) subunits, respectively. Conventionally, sucrose density gradient ultracentrifugation and SEC have been used to separate these capsids. However, good separation of the large and small particles with these methods is never achieved. In the present study, we employed a simple, fast, and cost-effective method to separate the T = 3 and T = 4 HBcAg capsids by using native agarose gel electrophoresis followed by an electroelution method (NAGE-EE). This is a direct, fast, and economic method for isolating the large and small HBcAg particles homogenously based on the hydrodynamic radius of the spherical particles. Dynamic light scattering analysis demonstrated that the T = 3 and T = 4 HBcAg capsids prepared using the NAGE-EE method are monodisperse with polydispersity values of ∼15% and ∼13%, respectively. ELISA proved that the antigenicity of the capsids was not affected in the purification process. Overall, NAGE-EE produced T = 3 and T = 4 capsids with a purity above 90%, and the recovery was 34% and 50%, respectively (total recovery of HBcAg is ∼84%), and the operation time is 15 and 4 times lesser than that of the sucrose density gradient ultracentrifugation and SEC, respectively.

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Display of the antigenic region of Nipah virus nucleocapsid protein on hepatitis B virus capsid

Cited by 9 publications

References 11 publications

Induction of Humoral and Cell-Mediated Immune Responses by Hepatitis B Virus Epitope Displayed on the Virus-Like Particles of Prawn Nodavirus

Induction of Humoral and Cell-Mediated Immune Responses by Hepatitis B Virus Epitope Displayed on the Virus-Like Particles of Prawn Nodavirus

Advances in the study of nodavirus

Native agarose gel electrophoresis and electroelution: A fast and cost‐effective method to separate the small and large hepatitis B capsids

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