Construction and immunogenicity of a recombinant pseudotype baculovirus expressing the glycoprotein of rabies virus in mice

Huang, Huaiyang; Xiao, Shaobo; Qin, Junling; Jiang, Yunbo; Wang, Cuiling; Li, Tingting; Gao, Yuwei; Li, Zilong; Li, Tiansong; Su, Xiuchan; Ruan, Yang; Xu, Fengqin; Wang, Hualei; Chen, Huanchun; Xia, Xianzhu

doi:10.1007/s00705-010-0909-4

Cited by 12 publications

(12 citation statements)

References 45 publications

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“…The baculovirus vector has several advantages when compared to other viral vectors as it can potentiate innate immune response against viral infections [1]. Also, the pseudotyped baculovirus can trigger a protective immune response in mice models [4,9]. Considering these promising results, it is desirable to effectively harness the potential of pseudotyped baculovirus as a viral vector for vaccine antigen delivery.…”

Section: Discussionmentioning

confidence: 99%

“…However, several researchers [9,19] have reported the utility of vesicular stomatitis virus glycoprotein (VSV-G, 511 amino acids long) pseudotyping of baculovirus for gene delivery into mammalian systems. Furthermore, while some researchers have utilized fulllength VSV-G for pseudotyping of baculovirus [9,19], there are also reports that truncated VSV-G, termed as VSV-GED, having a 21-amino-acid G-stem of ectodomain in conjunction with the transmembrane (TM) and C-terminal (CT) domain, was sufficient and increased the transduction efficiency of baculovirus-mediated gene delivery [4,11]. Thus far, there is no systematic comparative study on the identical platform investigating if VSV-G pseudotyping of the baculovirus is redundant given that the virus already has a similar functional protein, GP64.…”

Section: Introductionmentioning

confidence: 99%

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Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

et al. 2014

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The recombinant baculoviruses were constructed to investigate the necessity of VSV-G pseudotyping for mammalian cell transduction. The viruses were designed to express green fluorescent protein (GFP) gene under the control of cytomegalovirus promoter, with or without pseudotyping with VSV-G. VSV-G was placed under the control of polyhedrin promoter that is recognized by insect cells, allowing the formation of pseudotyped baculovirus. The study findings demonstrate that the pseudotyping of baculovirus significantly enhanced transduction efficiency compared to non-pseudotyped baculovirus, resulting in consequent distinction in the expression of GFP in mammalian cells. The results confirmed that pseudotyping is important for baculovirus mediated gene delivery. Further, when full-length VSV-G pseudotyping was compared with truncated VSV-G containing GED domain (G-stem of ectodomain in conjunction with the TM and CT domains of the glycoprotein), latter was relatively less efficient in transducing mammalian cells. This study demonstrated that pseudotyping with full-length VSV-G had better transduction efficiency in mammalian cells. However, at higher multiplicity of infection, both fulllength and truncated VSV-G showed equivalent transduction. This study established the significance of pseudotyping of baculovirus with full-length VSV-G for efficient transduction of mammalian cells, utilizing the highly sensitive GFP marker system. These findings have significant implications in designing of baculovirus vector based antigen delivery for developing new generation vaccines.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

et al. 2014

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“…In raccoons, oral immunization with RVG-containing insect cell lysate induced RV-neutralizing antibody titers that protected most but not all of the animals from lethal challenge (5). A pseudotype baculovirus expressing the G of RV was found to be safe and immunogenic in mice (12). Since the use of cells or cell lysate as a vaccine may not be appropriate, we tried to extract the membrane-bound r-RVG using several combinations of salts and detergents.…”

Section: Discussionmentioning

confidence: 99%

Expression and Solubilization of Insect Cell-Based Rabies Virus Glycoprotein and Assessment of Its Immunogenicity and Protective Efficacy in Mice

Rajendran

Subramanian

Sivakumar

et al. 2011

Clin. Vaccine Immunol.

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Rabies is a fatal zoonotic disease of serious public health and economic significance worldwide. The rabies virus glycoprotein (RVG) has been the major target for subunit vaccine development, since it harbors domains responsible for induction of virus-neutralizing antibodies, infectivity, and neurovirulence. The glycoprotein (G) was cloned using the baculovirus expression vector system (BEVS) and expressed in Spodoptera frugiperda (Sf-9) cells. In order to obtain a soluble form of G suitable for experimentation in mice, 18 different combinations of buffers and detergents were evaluated for their ability to solubilize the insect cell membraneassociated G. The combination that involved 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) detergent in lysis buffer 1, formulated with Tris, NaCl, 10% dimethyl sulfoxide (DMSO), and EDTA, gave the highest yield of soluble G, as evidenced by the experimental data. Subsequently, several other parameters, such as the concentration of CHAPS and the duration and temperature of the treatment for the effective solubilization of G, were optimized. The CHAPS detergent, buffered at a concentration of 0.4% to 0.7% (wt/vol) at room temperature (23 to 25°C) for 30 min to 1 h using buffer 1, containing 10% DMSO, resulted in consistently high yields. The G solubilized using CHAPS detergent was found to be immunogenic when tested in mice, as evidenced by high virus-neutralizing antibody titers in sera and 100% protection upon virulent intracerebral challenge with the challenge virus standard (CVS) strain of rabies virus. The results of the mice study indicated that G solubilized with CHAPS detergent retained the immunologically relevant domains in the native conformation, thereby paving the way for producing a cell-free and efficacious subunit vaccine.

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“…The glycoprotein G of RABV (RABV-G) assembles in the membrane envelope of the virion in the form of homotrimers, which harbour the major antigenic determinants of the virus [4]. Therefore, most of the recombinant candidate vaccines described today, are based on RABV-G protein produced with several different expression systems [5][6][7][8][9]. Some of these candidates were shown to be immunogenic in mice, but none have so far been registered for human or animal use.…”

Section: Introductionmentioning

confidence: 99%

A recombinant rabies vaccine expressing the trimeric form of the glycoprotein confers enhanced immunogenicity and protection in outbred mice

Koraka

Bosch

Cox

et al. 2014

Vaccine

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Rabies is a disease characterized by an invariably lethal encephalitis of viral origin that can be controlled by preventive vaccination programs of wildlife, domestic animals and humans in areas with a high risk of exposure. Currently available vaccines are expensive, cumbersome to produce and require intensive immunization and booster schemes to induce and maintain protective immunity. In the present study, we describe the development of candidate recombinant subunit rabies vaccines based on the glycoprotein G of the prototype rabies virus (RABV-G) expressed either as a monomer (RABV-mG) or in its native trimeric configuration (RABV-tG), with or without Matrix-M™ adjuvant. Immunogenicity and protective efficacy of the respective candidate vaccines were tested in outbred NIH Swiss albino mice. The RABV-tG candidate vaccine proved to be superior to the RABV-mG vaccine candidate both in terms of immunogenicity and efficacy. The relatively poor immunogenicity of the RABV-mG vaccine candidate was greatly improved by the addition of the adjuvant. A single, low dose of RABV-tG in combination with Matrix-M™ induced high levels of high avidity neutralizing antibodies and protected all mice against challenge with a lethal dose of RABV. Consequently RABV-tG used in combination with Matrix-M™ is a promising vaccine candidate that overcomes the limitations of currently used vaccines.

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Construction and immunogenicity of a recombinant pseudotype baculovirus expressing the glycoprotein of rabies virus in mice

Cited by 12 publications

References 45 publications

Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

Expression and Solubilization of Insect Cell-Based Rabies Virus Glycoprotein and Assessment of Its Immunogenicity and Protective Efficacy in Mice

A recombinant rabies vaccine expressing the trimeric form of the glycoprotein confers enhanced immunogenicity and protection in outbred mice

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