Functional cooperativity mediated by rationally selected combinations of human monoclonal antibodies targeting the henipavirus receptor binding protein

AbstractHendra virus (HeV) and Nipah virus (NiV), the prototypic members of the Henipavirus (HNV) genus, are emerging, zoonotic paramyxoviruses known to cause severe disease across six mammalian orders, including humans (Eaton et al., 2006). While several research groups have made strides in developing candidate vaccines and therapeutics against henipaviruses, such countermeasures have not been licensed for human use, and significant gaps in knowledge about the human immune response to these viruses exist. To address these gaps, we isolated a large panel of human monoclonal antibodies (mAbs) from the B cells of an individual with prior occupation-related exposure to the equine HeV vaccine (Equivac® HeV). Competition-binding and hydrogen-deuterium exchange mass spectrometry (HDX-MS) studies identified at least six distinct antigenic sites on the HeV/NiV receptor binding protein (RBP) that are recognized by human mAbs. Antibodies recognizing multiple antigenic sites potently neutralized NiV and/or HeV isolates in vitro. The most potent class of cross-reactive antibodies achieved neutralization by blocking viral attachment to the host cell receptors ephrin-B2 and ephrin-B3. Antibodies from this class mimic receptor binding by inducing a receptor-bound conformation to the HeV-RBP protein tetramer, exposing an epitope that appears to lie hidden in the interface between protomers within the HeV-RBP tetramer. Antibodies that recognize this cryptic epitope potently neutralized HeV and NiV. Flow cytometric studies using cell-surface-displayed HeV-RBP protein showed that cross-reactive, neutralizing mAbs from each of these classes cooperate for binding. In a highly stringent hamster model of NiVB infection, antibodies from both classes reduced morbidity and mortality and achieved synergistic protection in combination and provided therapeutic benefit when combined into two bispecific platforms. These studies identified multiple candidate mAbs that might be suitable for use in a cocktail therapeutic approach to achieve synergistic antiviral potency and reduce the risk of virus escape during treatment.

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Cooperativity mediated by rationally selected combinations of human monoclonal antibodies targeting the henipavirus receptor binding protein

Cell Reports ◽

10.1016/j.celrep.2021.109628 ◽

2021 ◽

Vol 36 (9) ◽

pp. 109628

Author(s):

Michael P. Doyle ◽

Nurgun Kose ◽

Viktoriya Borisevich ◽

Elad Binshtein ◽

Moushimi Amaya ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Receptor Binding ◽

Binding Protein ◽

Human Monoclonal Antibodies ◽

Receptor Binding Protein

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Isolation of steroid receptor binding protein from chicken oviduct and production of monoclonal antibodies

Biochemistry ◽

10.1021/bi00336a060 ◽

1985 ◽

Vol 24 (15) ◽

pp. 4214-4222 ◽

Cited By ~ 74

Author(s):

William P. Sullivan ◽

Benjamin T. Vroman ◽

Vickie J. Bauer ◽

Raj K. Puri ◽

Robert M. Riehl ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Receptor Binding ◽

Binding Protein ◽

Steroid Receptor ◽

Chicken Oviduct ◽

Receptor Binding Protein

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Molecular and Biological Characterization of Human Monoclonal Antibodies Binding to the Spike and Nucleocapsid Proteins of Severe Acute Respiratory Syndrome Coronavirus

Journal of Virology ◽

10.1128/jvi.79.3.1635-1644.2005 ◽

2005 ◽

Vol 79 (3) ◽

pp. 1635-1644 ◽

Cited By ~ 81

Author(s):

Edward N. van den Brink ◽

Jan ter Meulen ◽

Freek Cox ◽

Mandy A. C. Jongeneelen ◽

Alexandra Thijsse ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Severe Acute Respiratory Syndrome ◽

Receptor Binding ◽

Vero Cells ◽

Human Monoclonal Antibodies ◽

Human Mabs ◽

N Protein ◽

Antibody Phage ◽

Antibody Phage Display

ABSTRACT Human monoclonal antibodies (MAbs) were selected from semisynthetic antibody phage display libraries by using whole irradiated severe acute respiratory syndrome (SARS) coronavirus (CoV) virions as target. We identified eight human MAbs binding to virus and infected cells, six of which could be mapped to two SARS-CoV structural proteins: the nucleocapsid (N) and spike (S) proteins. Two MAbs reacted with N protein. One of the N protein MAbs recognized a linear epitope conserved between all published human and animal SARS-CoV isolates, and the other bound to a nonlinear N epitope. These two N MAbs did not compete for binding to SARS-CoV. Four MAbs reacted with the S glycoprotein, and three of these MAbs neutralized SARS-CoV in vitro. All three neutralizing anti-S MAbs bound a recombinant S1 fragment comprising residues 318 to 510, a region previously identified as the SARS-CoV S receptor binding domain; the nonneutralizing MAb did not. Two strongly neutralizing anti-S1 MAbs blocked the binding of a recombinant S fragment (residues 1 to 565) to SARS-CoV-susceptible Vero cells completely, whereas a poorly neutralizing S1 MAb blocked binding only partially. The MAb ability to block S1-receptor binding and the level of neutralization of the two strongly neutralizing S1 MAbs correlated with the binding affinity to the S1 domain. Finally, epitope mapping, using recombinant S fragments (residues 318 to 510) containing naturally occurring mutations, revealed the importance of residue N479 for the binding of the most potent neutralizing MAb, CR3014. The complete set of SARS-CoV MAbs described here may be useful for diagnosis, chemoprophylaxis, and therapy of SARS-CoV infection and disease.

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Localization of the growth hormone receptor/binding protein in skin

Journal of Endocrinology ◽

10.1677/joe.0.1260467 ◽

1990 ◽

Vol 126 (3) ◽

pp. 467-NP ◽

Cited By ~ 60

Author(s):

P. E. Lobie ◽

W. Breipohl ◽

D. T. Lincoln ◽

J. García-Aragón ◽

M. J. Waters

Keyword(s):

Monoclonal Antibodies ◽

Receptor Binding ◽

Growth Hormone Receptor ◽

Binding Protein ◽

Cell Types ◽

Myoepithelial Cells ◽

Hair Follicles ◽

Sweat Glands ◽

Gh Receptor ◽

Receptor Binding Protein

ABSTRACT Acromegaly is characterized by coarsening of facial features, acanthosis nigricans, hypertrichosis and oily skin. To determine the site through which GH exerts these effects, we have used immunohistochemistry to localize the GH receptor/binding protein (BP) in rat, rabbit and human skin. Three monoclonal antibodies (MAb 1, 43, 263) were immunoreactive in identical locations, whereas no immunoreactivity was evident when control monoclonal antibodies (MAb 50.8 and MAb 7 (rat)) were used. Skin from neonatal and adult animals was used to determine whether GH receptor/BP expression was developmentally regulated. Immunoreactivity of the GH receptor/BP in the three species was consistently localized in the stratum basale and stratum spinosum. Intermittent staining was observed in the stratum granulosum. Scattered basal epidermal cells often displayed more intense immunoreactivity. This distribution was observed at all maturational stages examined. Intense GH receptor/BP immunoreactivity was observed in all histological layers of the lower one-third of hair follicles and in hair matrix cells of the dermal papillae. Immunoreactivity was also detected in the outer epithelial root sheath of the upper two-thirds of hair follicles, in sebaceous glands and in fibroblasts of the connective tissue sheath surrounding the follicle. GH receptor/BP immunoreactivity was also present in the secretory duct and myoepithelial cells of human eccrine sweat glands. Fibroblasts, Schwann cells of peripheral nerve fascicles, skeletal muscle cells and adipocytes of the dermis were also immunoreactive as were medial smooth muscle and endothelial cells of arteries. These results provide evidence that GH acts locally on the epidermis and epidermal appendages concordant with our recent localization of GH receptor/BP to epithelial cell types of the gastrointestinal and reproductive systems. Journal of Endocrinology (1990) 126, 467–472

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