title: G glycoprotein amino acid residues required for human monoclonal antibody RAB1 neutralization are conserved in rabies virus street isolates.
abstract: Replacement of polyclonal anti-rabies immunoglobulin (RIG) used in rabies post-exposure prophylaxis (PEP) with a monoclonal antibody will eliminate cost and availability constraints that currently exist using RIG in the developing world. The human monoclonal antibody RAB1 has been shown to neutralize all rabies street isolates tested; however for the laboratory-adapted fixed strain, CVS-11, mutation in the G glycoprotein of amino acid 336 from asparagine (N) to aspartic acid (D) resulted in resistance to neutralization. Interestingly, this same mutation in the G glycoprotein of a second laboratory-adapted fixed strain (ERA) did not confer resistance to RAB1 neutralization. Using cell surface staining and lentivirus pseudotyped with rabies virus G glycoprotein (RABVpp), we identified an amino acid alteration in CVS-11 (K346), not present in ERA (R346), which was required in combination with D336 to confer resistance to RAB1. A complete analysis of G glycoprotein sequences from GenBank demonstrated that no identified rabies isolates contain the necessary combination of G glycoprotein mutations for resistance to RAB1 neutralization, consistent with the broad neutralization of RAB1 observed in direct viral neutralization experiments with street isolates. All combinations of amino acids 336 and 346 reported in the sequence database were engineered into the ERA G glycoprotein and RAB1 was able to neutralize RABVpp bearing ERA G glycoprotein containing all known combinations at these critical residues. These data demonstrate that RAB1 has the capacity to neutralize all identified rabies isolates and a minimum of two distinct mutations in the G glycoprotein are required for abrogation of RAB1 neutralization.
authors.affiliation: MassBiologics, University of Massachusetts Medical School, Boston, MA 02126, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Kirk J
authors.initials: KJ
authors.lastname: Rowley
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Booth
authors.forename: Susan E
authors.initials: SE
authors.lastname: Sloan
authors.forename: Donna M
authors.initials: DM
authors.lastname: Ambrosino
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Babcock
mesh_terms: D019943:Amino Acid Substitution; D000911:Antibodies, Monoclonal / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001216:Asparagine / Q000378:metabolism; D001666:Binding Sites, Antibody; D003001:Cloning, Molecular; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016297:Mutagenesis, Site-Directed / Q000379:methods; D009500:Neutralization Tests; D017354:Point Mutation; D011820:Rabies virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020539:Sequence Analysis, Protein; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus; D001216:Asparagine