Immunogenic, non-infectious polio subviral particles synthesized in Saccharomyces cerevisiae.

Rombaut, Bartholomeus; Jore, J.p.m.

doi:10.1099/0022-1317-78-8-1829

Cited by 27 publications

(16 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immunogenic, but unstable, VLPs were also produced by recombinant expression in yeast [16]. A strategy to address this instability is described here where candidate stabilising mutations were identified then multiple changes introduced into capsid proteins in combination.…”

Section: Discussionmentioning

confidence: 99%

Genetically Thermo-Stabilised, Immunogenic Poliovirus Empty Capsids; a Strategy for Non-replicating Vaccines

Fox¹,

Knowlson²,

Minor³

et al. 2017

PLoS Pathog

View full text Add to dashboard Cite

While wild type polio has been nearly eradicated there will be a need to continue immunisation programmes for some time because of the possibility of re-emergence and the existence of long term excreters of poliovirus. All vaccines in current use depend on growth of virus and most of the non-replicating (inactivated) vaccines involve wild type viruses known to cause poliomyelitis. The attenuated vaccine strains involved in the eradication programme have been used to develop new inactivated vaccines as production is thought safer. However it is known that the Sabin vaccine strains are genetically unstable and can revert to a virulent transmissible form. A possible solution to the need for virus growth would be to generate empty viral capsids by recombinant technology, but hitherto such particles are so unstable as to be unusable. We report here the genetic manipulation of the virus to generate stable empty capsids for all three serotypes. The particles are shown to be extremely stable and to generate high levels of protective antibodies in animal models.

show abstract

Section: Discussionmentioning

confidence: 99%

Genetically Thermo-Stabilised, Immunogenic Poliovirus Empty Capsids; a Strategy for Non-replicating Vaccines

Fox¹,

Knowlson²,

Minor³

et al. 2017

PLoS Pathog

View full text Add to dashboard Cite

show abstract

“…To our knowledge, this is the first time that this has been reported for SVDV, although it has been done for other enteroviruses, by use of baculovirus recombinants in insect cells (6,21,39), vaccinia recombinants in mammalian cells (2), and the yeast Saccharomyces cerevisiae (35). Recombinant VLPs have been produced on the basis of the expression of full-length polyprotein (39), the P1 and 3CD proteins (21,35), the P1 and P3 proteins (2), or the individual VP0, VP3, and VP1 proteins (6) in several expression systems. The yield (20 to 180 g/liter) of VLP obtained by single recombinant (based on the expression of full-length polyprotein) (39) was enhanced compared to that obtained by multiple baculovirus recombinants (6, 21) but was still not satisfactory for development of a subunit vaccine or diagnostic antigen.…”

Section: Discussionmentioning

confidence: 99%

Noninfectious Virus-Like Particle Antigen for Detection of Swine Vesicular Disease Virus Antibodies in Pigs by Enzyme-Linked Immunosorbent Assay

Choi

Nah

et al. 2005

Clin Vaccine Immunol

View full text Add to dashboard Cite

An inactivated SVDV antigen is used in current enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies to swine vesicular disease virus (SVDV). To develop a noninfectious recombinant alternative, we produced SVDV-like particles (VLPs) morphologically and antigenically resembling authentic SVDV particles by using a dual baculovirus recombinant, which expresses simultaneously the P1 and 3CD protein genes of SVDV under different promoters. Antigenic differences between recombinant VLPs and SVDV particles were not statistically significant in results obtained with a 5B7-ELISA kit, indicating that the VLPs could be used in the place of SVDV antigen in ELISA kits. We developed a blocking ELISA using the VLPs and SVDV-specific neutralizing monoclonal antibody 3H10 (VLP-ELISA) for detection of SVDV serum antibodies in pigs. The VLP-ELISA showed a high specificity of 99.9% when tested with pig sera that are negative for SVDV neutralization (n ‫؍‬ 1,041). When tested using sera (n ‫؍‬ 186) collected periodically from pigs (n ‫؍‬ 19) with experimental infection with each of three different strains of SVDV, the VLP-ELISA detected SVDV serum antibodies as early as 3 days postinfection and continued to detect the antibodies from all infected pigs until termination of the experiments (up to 121 days postinfection). This test performance was similar to that of the gold standard virus neutralization test and indicates that the VLP-ELISA is a highly specific and sensitive method for the detection of SVDV serum antibodies in pigs. This is the first report of the production and diagnostic application of recombinant VLPs of SVDV. Further potential uses of the VLPs are discussed.

show abstract

“…Earlier studies demonstrated the production of empty capsids of several picornaviruses following translation of the viral RNA in rabbit reticulocyte lysates (25,30,45) or expression of capsid precursor in bacterial (34), yeast (55), insect (14) and mammalian (1,3) cells. In these studies, capsid precursors were coexpressed with the viral protease in order for capsid assembly to occur within the expression system.…”

mentioning

confidence: 99%

Foot-and-Mouth Disease Virus Assembly: Processing of Recombinant Capsid Precursor by Exogenous Protease Induces Self-Assembly of Pentamers In Vitro in a Myristoylation-Dependent Manner

Goodwin

Tuthill

Arias

et al. 2009

J Virol

View full text Add to dashboard Cite

The assembly of foot-and-mouth disease virus (FMDV) particles is poorly understood. In addition, there are important differences in the antigenic and receptor binding properties of virus assembly and dissociation intermediates, and these also remain unexplained. We have established an experimental model in which the antigenicity, receptor binding characteristics, and in vitro assembly of capsid precursor can be studied entirely from purified components. Recombinant capsid precursor protein (P1 region) was expressed in Escherichia coli as myristoylated or unmyristoylated protein. The protein sedimented in sucrose gradients at 5S and reacted with monoclonal antibodies which recognize conformational or linear antigen determinants on the virion surface. In addition, it bound the integrin ␣ v ␤ 6 , a cellular receptor for FMDV, indicating that unprocessed recombinant capsid precursor is both structurally and antigenically similar to native virus capsid. These characteristics were not dependent on the presence of 2A at the C terminus but were altered by N-terminal myristoylation and in mutant precursors which lacked VP4. Proteolytic processing of myristoylated precursor by recombinant FMDV 3C pro in vitro induced a shift in sedimentation from 5S to 12S, indicating assembly into pentameric capsid subunits. Nonmyristoylated precursor still assembled into higher-order structures after processing with 3Cpro , but these particles sedimented in sucrose gradients at approximately 17S. In contrast, mutant precursors lacking VP4 were antigenically distinct, were unable to form pentamers, and had reduced capacity for binding integrin receptor. These studies demonstrate the utility of recombinant capsid precursor protein for investigating the initial stages of assembly of FMDV and other picornaviruses.Foot-and-mouth disease virus (FMDV) is the etiological agent of a highly infectious disease of cattle and other clovenhoofed animals. It is of economic importance, because the presence of the disease results in severe restrictions of international trade. In many parts of the developing world, the disease is endemic, and it continues to pose a serious threat to livestock industries globally, as exemplified by recent major outbreaks in the United Kingdom, Argentina, and Uruguay.FMDV is a small, nonenveloped, positive-strand RNA virus belonging to the genus Aphthovirus within the family Picornaviridae. Other members of the family include poliovirus (PV; genus Enterovirus) and hepatitis A virus (genus Hepatovirus). The mature picornavirus comprises 60 copies each of four structural proteins, termed VP1, VP2, VP3, and VP4, which encapsidate a single, positive-sense RNA genome. The proteins form a pseudo Tϭ3 icosahedral capsid with VP1 located close to the fivefold axes of symmetry and VP2 and VP3 alternating around the threefold axes (24). VP4, which is myristoylated at the N terminus, is an internal component of the capsid (13,48).Although the morphogenesis of picornaviral particles is not completely understood, it is known that capsid...

show abstract

Immunogenic, non-infectious polio subviral particles synthesized in Saccharomyces cerevisiae.

Cited by 27 publications

References 14 publications

Genetically Thermo-Stabilised, Immunogenic Poliovirus Empty Capsids; a Strategy for Non-replicating Vaccines

Genetically Thermo-Stabilised, Immunogenic Poliovirus Empty Capsids; a Strategy for Non-replicating Vaccines

Noninfectious Virus-Like Particle Antigen for Detection of Swine Vesicular Disease Virus Antibodies in Pigs by Enzyme-Linked Immunosorbent Assay

Foot-and-Mouth Disease Virus Assembly: Processing of Recombinant Capsid Precursor by Exogenous Protease Induces Self-Assembly of Pentamers In Vitro in a Myristoylation-Dependent Manner

Contact Info

Product

Resources

About