title: Human Memory B Cells Producing Potent Cross-Neutralizing Antibodies against Human Parechovirus: Implications for Prevalence, Treatment, and Diagnosis.
abstract: The family Picornaviridae is a large and diverse group of positive-sense RNA viruses, including human enteroviruses (EVs) and human parechoviruses (HPeVs). The human immune response against EVs and HPeVs is thought to be mainly humoral, and an insufficient neutralizing antibody (Ab) response during infection is a risk factor and can ultimately be life threatening. The accessibility of different antigenic sites and observed cross-reactivity make HPeVs a good target for development of therapeutic human monoclonal antibodies (MAbs). In this study, we generated two different human MAbs specific for HPeV by screening culture supernatants of Ab-producing human B cell cultures for direct neutralization of HPeV1. Both MAbs showed HPeV1-specific neutralization as well as neutralization of HPeV2. One antibody, AM18, cross-neutralized HPeV4, -5, and -6 and coxsackievirus A9 (CV-A9). VP1 capsid protein-specific assays confirmed that AM18 bound VP1 of HPeV1, -2, and -4 with high affinity (11.5 pM). In contrast, the HPeV1-specific MAb AM28, which neutralized HPeV1 even more efficiently than did AM18, showed no cross-reactivity with HPeV3 to -6 or other EVs and did not bind any of the capsid proteins, suggesting that AM28 is specific for a conformation-dependent, nonlinear epitope on the virus. The discovery of MAbs that are cross-reactive between HPeVs may help development of HPeV treatment options with antibodies and vaccine design based on epitopes recognized by these antibodies.
  
  
  OBJECTIVE
  HPeV infections are widespread among young children and adults, causing a broad range of disease. Infections can be severe and life threatening, while no antiviral treatment is available. Given that the absence of neutralizing Abs is a risk factor for severe disease in infants, treatment of picornavirus infections with MAbs would be a therapeutic option. To study antibody neutralization of HPeV in more detail, we generated two different HPeV1-specific human MAbs. Both MAbs show HPeV1-specific neutralization and cross-neutralized HPeV2. One MAb also cross-neutralized other HPeVs. Surprisingly, this MAb also neutralized CV-A9. These MAbs provide a unique tool for further research and for the diagnosis (antigen detection) and possible treatment of HPeV infections.
authors.affiliation: Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands westerhuis.brenda@gmail.com beaumont@aimmtherapeutics.com.
authors.forename: B M
authors.initials: BM
authors.lastname: Westerhuis
authors.affiliation: Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: K S M
authors.initials: KS
authors.lastname: Benschop
authors.affiliation: Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: G
authors.initials: G
authors.lastname: Koen
authors.affiliation: AIMM Therapeutics, Academic Medical Center, Amsterdam, the Netherlands.
authors.forename: Y B
authors.initials: YB
authors.lastname: Claassen
authors.affiliation: AIMM Therapeutics, Academic Medical Center, Amsterdam, the Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Wagner
authors.affiliation: AIMM Therapeutics, Academic Medical Center, Amsterdam, the Netherlands.
authors.forename: A Q
authors.initials: AQ
authors.lastname: Bakker
authors.affiliation: Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: K C
authors.initials: KC
authors.lastname: Wolthers
authors.affiliation: AIMM Therapeutics, Academic Medical Center, Amsterdam, the Netherlands westerhuis.brenda@gmail.com beaumont@aimmtherapeutics.com.
authors.forename: T
authors.initials: T
authors.lastname: Beaumont
mesh_terms: D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology* / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology; D003429:Cross Reactions; D006801:Humans; D009426:Netherlands / Q000453:epidemiology; D030061:Parechovirus / Q000145:classification / Q000235:genetics / Q000276:immunology*; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000276:immunology* / Q000628:therapy; D015995:Prevalence
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D036022:Capsid Proteins