Human Memory B Cells Producing Potent Cross-Neutralizing Antibodies against Human Parechovirus: Implications for Prevalence, Treatment, and Diagnosis

Westerhuis, Brenda M.; Benschop, Kimberley; Koen, Gerrit; Claassen, Yvonne; Wagner, Kay Uwe; Bakker, Arjen Q.; Wolthers, Katja C.; Beaumont, Tim

doi:10.1128/jvi.01079-15

Cited by 23 publications

(32 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Two antibodies (AM18 and AM28) were selected, and the corresponding nucleotide sequences were cloned for expression in 293T cells and purified. The initial production and characterization of these MAbs have been recently described in detail (21). Initially, we tested binding of each MAb to HPeV1 virions at molar ratios of 60:1 and 300:1 at 37°C for 1 h in 1ϫ TNM buffer followed by cryo-EM (see below).…”

Section: Methodsmentioning

confidence: 99%

“…Human memory IgG ϩ B cells were cultured using the AIMSelect method (23), and antibody-containing culture supernatants were used to directly screen for HPeV1 neutralization (21). Two antibodies (AM18 and AM28) were selected, and the corresponding nucleotide sequences were cloned for expression in 293T cells and purified.…”

Section: Methodsmentioning

confidence: 99%

“…We have isolated two different human HPeV1 monoclonal antibodies (MAbs), of which MAb AM18 was shown to be a broadly cross-neutralizing MAb against HPeV1, -2, -4, -5, and -6 and MAb AM28 neutralized HPeV1 and HPeV2 (21). These results indicated two different neutralizing epitopes for AM18 and AM28.…”

mentioning

confidence: 97%

See 2 more Smart Citations

Structural Basis of Human Parechovirus Neutralization by Human Monoclonal Antibodies

Shakeel

Westerhuis

Ora

et al. 2015

J Virol

View full text Add to dashboard Cite

Since it was first recognized in 2004 that human parechoviruses (HPeV) are a significant cause of central nervous system and neonatal sepsis, their clinical importance, primarily in children, has started to emerge. Intravenous immunoglobulin treatment is the only treatment available in such life-threatening cases and has given moderate success. Direct inhibition of parechovirus infection using monoclonal antibodies is a potential treatment. We have developed two neutralizing monoclonal antibodies against HPeV1 and HPeV2, namely, AM18 and AM28, which also cross-neutralize other viruses. Here, we present the mapping of their epitopes using peptide scanning, surface plasmon resonance, fluorescence-based thermal shift assays, electron cryomicroscopy, and image reconstruction. We determined by peptide scanning and surface plasmon resonance that AM18 recognizes a linear epitope motif including the arginine-glycine-aspartic acid on the C terminus of capsid protein VP1. This epitope is normally used by the virus to attach to host cell surface integrins during entry and is found in 3 other viruses that AM18 neutralizes. Therefore, AM18 is likely to cause virus neutralization by aggregation and by blocking integrin binding to the capsid. Further, we show by electron cryomicroscopy, three-dimensional reconstruction, and pseudoatomic model fitting that ordered RNA interacts with HPeV1 VP1 and VP3. AM28 recognizes quaternary epitopes on the capsid composed of VP0 and VP3 loops from neighboring pentamers, thereby increasing the RNA accessibility temperature for the virus-AM28 complex compared to the virus alone. Thus, inhibition of RNA uncoating probably contributes to neutralization by AM28. IMPORTANCEHuman parechoviruses can cause mild infections to severe diseases in young children, such as neonatal sepsis, encephalitis, and cardiomyopathy. Intravenous immunoglobulin treatment is the only treatment available in such life-threatening cases. In order to develop more targeted treatment, we have searched for human monoclonal antibodies that would neutralize human parechoviruses 1 and 2, associated with mild infections such as gastroenteritis and severe infections of the central nervous system, and thus allow safe treatment. In the current study, we show how two such promising antibodies interact with the virus, modeling the atomic interactions between the virus and the antibody to propose how neutralization occurs. Both antibodies can cause aggregation; in addition, one antibody interferes with the virus recognizing its target cell, while the other, recognizing only the whole virus, inhibits the genome uncoating and replication in the cell. Human parechoviruses (HPeV) are single-stranded, positivesense RNA viruses in the Parechovirus genus within the Picornaviridae family (1). HPeV infections mainly cause mild gastrointestinal symptoms, although HPeV are also associated with more severe central nervous system symptoms, such as meningitis and neonatal sepsis (2-5). The HPeV genome is about 7,300 bases in length, enclose...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Structural Basis of Human Parechovirus Neutralization by Human Monoclonal Antibodies

Shakeel

Westerhuis

Ora

et al. 2015

J Virol

View full text Add to dashboard Cite

show abstract

“…One is localized to residues 83-97 of VP0 [172], another is found on VP1 and encompasses the receptor-recognizing arginine-glycine-aspartic acid (RGD) motif [173], and the third one is formed by VP0 and VP3 [169,174]. Whereas antisera generated against the HPeV-1 VP0 peptide were not tested for HPeV-3 neutralization, two other monoclonal antibodies did not cross-react with HPeV-3 [169,174]. Only a non-neutralizing epitope has been described for HPeV-3 [170].…”

Section: Neutralization Of Hpev-3mentioning

confidence: 99%

“…A useful approach for this is identification of an individual's immune response to a given virus followed by respective B-cell cloning [182]. This approach was utilized to generate two broadly neutralizing antibodies against HPeV [169,174]. Unfortunately, these antibodies did not neutralize HPeV-3 in a conventional microneutralization test.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Human picornaviruses associated with neurological diseases and their neutralization by antibodies

Anastasina

Domanska

Palm

et al. 2017

Journal of General Virology

View full text Add to dashboard Cite

Picornaviruses are the most commonly encountered infectious agents in mankind. They typically cause mild infections of the gastrointestinal or respiratory tract, but sometimes also invade the central nervous system. There, they can cause severe diseases with long-term sequelae and even be lethal. The most infamous picornavirus is poliovirus, for which significant epidemics of poliomyelitis were reported from the end of the nineteenth century. A successful vaccination campaign has brought poliovirus close to eradication, but neurological diseases caused by other picornaviruses have increasingly been reported since the late 1990s. In this review we focus on enterovirus 71, coxsackievirus A16, enterovirus 68 and human parechovirus 3, which have recently drawn attention because of their links to severe neurological diseases. We discuss the clinical relevance of these viruses and the primary role of humoral immunity in controlling them, and summarize current knowledge on the neutralization of such viruses by antibodies.

show abstract

Human Parechoviruses: Biology and Infection

Fischer

Harvala

Midgley

2017

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Infections with human parechoviruses (HPeVs) are highly prevalent, particularly in neonates, where they may cause substantial morbidity and mortality. The clinical presentation of HPeV infection in neonates is indistinguishable from that of enterovirus (EV) infection and may vary from mild disease involving gastrointestinal and/or respiratory symptoms to more severe illness with sepsis‐like symptoms such as fever, seizures, irritability, rash and/or feeding problems. HPeV is among the most common single causes of aseptic meningitis/meningoencephalitis in infants <90 days old worldwide. The mainstay of diagnosis for HPeV infections involves the use of molecular diagnostic techniques for detection of virus RNA (ribonucleic acid) in various materials (e.g. faeces, cerebrospinal fluid (CSF), nasopharyngeal, bronchial and autopsy materials). Despite the relatedness of HPeV and EV both at the genomic and clinical presentation levels, routine nucleic acid detection methods for EV will not detect HPeV. Laboratory diagnosis of these viral infections is important not only for differential diagnostic purposes and determining a patient's prognosis but also for guidance of clinical management. Clinical disease varies somewhat among genotypes; while HPeV1 mostly causes gastrointestinal illness, HPeV3 prominence is due to sepsis syndromes and central nervous system (CNS) infections in preterm and term infants. Of diagnostic importance, HPeV3 CNS infections usually lack CSF pleocytosis, and despite ‘unremarkable’ CSF, HPeV3 RNA can be amplified from CSF, nasopharyngeal and rectal swabs in an infected individual. Taking the global incidence of reported laboratory‐confirmed HPeV cases into consideration, mortality and sequelae are overall uncommon and usually accompanying initially severe or neurologically complicated acute illnesses. However, underreporting due to lack of testing and/or appropriate diagnostic methodology likely means gross underestimation of the true incidence of HPeV disease. Currently, neither effective treatment nor vaccines are available in the control of these common viruses with the potential to cause severe harm in neonates and young children worldwide. Key Concepts HPeV infections are common among infants and children worldwide. HPeV infections are underdiagnosed, and HPeV disease burden underestimated. Nineteen HPeV types identified so far – their role/significance in human infections unclear except for HPeV3. HPeV3 is a major cause of neonatal infections including severe meningoencephalitis. Genomic differences between EV and HPeV. Diagnostics: HPeV required specific PCR for detection, not that easy to culture. Majority of children with HPeV3 infection lack pleocytosis in CSF/have normal CSF. Having older siblings are associated with higher risk for an infant to be hospitalised with severe HPeV infection. HPeV infection should always be ruled out when caring for infants with sepsis‐like illness and/or CNS infection.

show abstract

Human Memory B Cells Producing Potent Cross-Neutralizing Antibodies against Human Parechovirus: Implications for Prevalence, Treatment, and Diagnosis

Cited by 23 publications

References 54 publications

Structural Basis of Human Parechovirus Neutralization by Human Monoclonal Antibodies

Structural Basis of Human Parechovirus Neutralization by Human Monoclonal Antibodies

Human picornaviruses associated with neurological diseases and their neutralization by antibodies

Human Parechoviruses: Biology and Infection

Contact Info

Product

Resources

About