The conserved N-terminus of human rhinovirus capsid protein VP4 contains membrane pore-forming activity and is a target for neutralizing antibodies

Panjwani, Anusha; Asfor, Amin S.; Tuthill, Tobias J.

doi:10.1099/jgv.0.000629

Cited by 22 publications

(41 citation statements)

References 21 publications

(30 reference statements)

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“…The obtained results agree with earlier results obtained with virus escape mutants as well as with virus-specific monoclonal and polyclonal antibodies, indicating that VP1 and VP2 contain epitopes involved in the RV–receptor interaction [ 28 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 ]. However, we could not show that VP3- or VP4-specific antibodies could block the RV–receptor interaction, although results from virus escape mutants [ 47 ] and studies performed with specific antibodies [ 55 , 56 , 57 , 58 ] indicated that VP3 and VP4, respectively, may play a role in virus neutralization. This discrepancy may be explained by VP3 and VP4 not being directly involved in receptor binding but may contribute to virus neutralization via other mechanisms.…”

Section: Discussionmentioning

confidence: 60%

ELISA-Based Assay for Studying Major and Minor Group Rhinovirus–Receptor Interactions

et al. 2020

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Rhinovirus (RV) infections are a major cause of recurrent common colds and trigger severe exacerbations of chronic respiratory diseases. Major challenges for the development of vaccines for RV include the virus occurring in the form of approximately 160 different serotypes, using different receptors, and the need for preclinical models for the screening of vaccine candidates and antiviral compounds. We report the establishment and characterization of an ELISA-based assay for studying major and minor group RV–receptor interactions. This assay is based on the interaction of purified virus with plate-bound human receptor proteins, intercellular adhesion molecule 1 (ICAM-1), and low density lipoprotein receptor (LDLR). Using RV strain-specific antibodies, we demonstrate the specific binding of a panel of major and minor RV group types including RV-A and RV-B strains to ICAM-1 and LDLR, respectively. We show that the RV–receptor interaction can be blocked with receptor-specific antibodies as well as with soluble receptors and neutralizing RV-specific antibodies. The assay is more sensitive than a cell culture-based virus neutralization test. The ELISA assay will therefore be useful for the preclinical evaluation for preventive and therapeutic strategies targeting the RV–receptor interaction, such as vaccines, antibodies, and anti-viral compounds.

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Section: Discussionmentioning

confidence: 60%

ELISA-Based Assay for Studying Major and Minor Group Rhinovirus–Receptor Interactions

et al. 2020

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“…VP2N45 was used for the development of the peptide ELISA. A control peptide equivalent to a capsid sequence from the related picornavirus human rhinovirus was used [18]. Eight peptides (15mer each) overlapping by ten amino acids, covering the first 45 amino acids from the N-terminus of the FMDV capsid sequence, were used for the fine mapping of the epitope (Fig.1a).…”

Section: Methodsmentioning

confidence: 99%

Detection of antibodies against a conserved capsid epitope as the basis of a novel universal serological test for foot-and-mouth disease

Asfor

Howe

Grazioli

et al. 2019

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16Diagnostic tests for foot-and-mouth disease (FMD) include the detection of antibodies 17 against either the viral non-structural proteins or the capsid. The detection of antibodies against 18 the structural proteins (SP) of the capsid can be used to monitor seroconversion in both infected 19 and vaccinated animals. However, SP tests need to be tailored to the individual FMD virus 20 serotype and their sensitivity performances may be affected by antigenic variability within each 21 serotype and mismatching between tests reagents. As a consequence, FMD Reference 22 Laboratories need to maintain contingency to employ multiple type-specific assays for large-23 scale serological surveillance and post-vaccination monitoring in the event of FMD outbreaks. 24 In this study, a highly conserved region in the N terminus of FMDV capsid protein VP2 (VP2N) 25 was characterised using a panel of intertypic-reactive monoclonal antibodies. This revealed a 26 universal epitope in VP2N which could be used as a peptide antigen to detect FMDV-specific 27 antibodies against all types of the virus. A VP2-peptide ELISA (VP2-ELISA) was optimised 28 using experimental and reference antisera from immunized, convalescent and negative animals 29 (n=172). The VP2-ELISA is universal, simple and provided sensitive (98.6 %) and specific 30 (93%) detection of antibodies to all FMDV strains used in this study. We anticipate that this 31 SP test could have utility for sero-surveillance during virus incursions in FMD-free countries 32 and as an additional screening tool to assess FMD virus circulation in endemic countries. 33 34 Keywords: FMDV, conserved capsid epitope, ELISA, diagnosis, serology. 35 36 37 Foot-and-mouth disease (FMD) is an economically devastating viral disease of cloven-38 hoofed animals with a global distribution. It limits access to markets for developing countries 39 and outbreaks in otherwise FMD-free countries are expensive to control (as in the UK in 2001, 40 Japan in 2010 and the Republic of Korea in 2010 and 2011) [1, 2]. FMD virus (FMDV) is a 41 single-stranded, positive-sense, RNA virus belonging to the genus Aphthovirus in the family 42 Picornaviridae. The virus exists as seven serotypes (O, A, C, Asia 1, South African Territories 43 (SAT)1, SAT2 and SAT3) as well as numerous and constantly evolving strains showing a 44 spectrum of antigenic diversity. 45The non-enveloped picornavirus capsid has icosahedral symmetry, a diameter of 46 approximately 30 nm and is composed of 60 copies of each of the capsid proteins VP1, VP2, 47 VP3 and VP4. VP1, VP2 and VP3 are the major components of the capsid, while VP4 is a 48 small (approximately (12 kDa) internal protein which lies on the inside surface of the capsid 49 around the five-fold axes of symmetry, where it is thought to stabilise interactions between 50 pentameric capsid subunits [3, 4]. During the replication cycle of FMDV, eight different viral 51 non-structural proteins (NSPs; and additional precursors) are generated which are potential 52 serological...

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“…A control peptide equivalent to a capsid sequence from the related picornavirus human rhinovirus was used [18]. Eight peptides (15mer each) overlapping by ten amino acids , covering the first 45 amino acids from the N-terminus of the FMDV capsid sequence, were used for the fine mapping of the epitope (Fig.1a).…”

Section: Peptidesmentioning

confidence: 99%

Detection of Bovine Antibodies against a Conserved Capsid Epitope as the Basis of a Novel Universal Serological Test for Foot-and-Mouth Disease

et al. 2020

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Diagnostic tests for foot-and-mouth disease (FMD) include the detection of antibodies against either the viral nonstructural proteins or the capsid. The detection of antibodies against the structural proteins (SP) of the capsid can be used to monitor seroconversion in both infected and vaccinated animals. However, SP tests need to be tailored to the individual FMD virus (FMDV) serotype and their sensitivity may be affected by antigenic variability within each serotype and mismatching between test reagents. As a consequence, FMD reference laboratories are required to maintain multiple type-specific SP assays and reagents. A universal SP test would simplify frontline diagnostics and facilitate large-scale serological surveillance and postvaccination monitoring. In this study, a highly conserved region in the N terminus of FMDV capsid protein VP2 (VP2N) was characterized using a panel of intertype-reactive monoclonal antibodies. This revealed a universal epitope in VP2N which could be used as a peptide antigen to detect FMDV-specific antibodies against all types of the virus. A VP2-peptide enzyme-linked immunosorbent assay (VP2-ELISA) was optimized using experimental and reference antisera from immunized, convalescent, and naïve animals (n = 172). The VP2-ELISA is universal and simple and provided sensitive (99%) and specific (93%) detection of antibodies to all FMDV strains used in this study. We anticipate that this SP test could have utility for serosurveillance during virus incursions in FMD-free countries and as an additional screening tool to assess FMD virus circulation in countries where the disease is endemic.

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The conserved N-terminus of human rhinovirus capsid protein VP4 contains membrane pore-forming activity and is a target for neutralizing antibodies

Cited by 22 publications

References 21 publications

ELISA-Based Assay for Studying Major and Minor Group Rhinovirus–Receptor Interactions

ELISA-Based Assay for Studying Major and Minor Group Rhinovirus–Receptor Interactions

Detection of antibodies against a conserved capsid epitope as the basis of a novel universal serological test for foot-and-mouth disease

Detection of Bovine Antibodies against a Conserved Capsid Epitope as the Basis of a Novel Universal Serological Test for Foot-and-Mouth Disease

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