Antibodies and methods for the diagnosis and treatment of Epstein Barr Virus infection
Inventors
Bu, Wei • Kanekiyo, Masaru • Joyce, Michael Gordon • Cohen, Jeffrey I.
Assignees
Henry M Jackson Foundation for Advancedment of Military Medicine Inc • US Department of Health and Human Services
Publication Number
US-11926656-B2
Publication Date
2024-03-12
Expiration Date
2038-04-25
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Abstract
Antibodies and compositions of matter useful for the detection, diagnosis and treatment of Epstein Barr Virus (EBV) infection in mammals, and methods of using those compositions of matter for the detection, diagnosis and treatment of EBV infection are described. Also described are proteins, referred to as anti-gp350 antibody probes, and anti-gp350 B-cell probes, that maintain the epitope structure of the CR2-binding region of gp350, but do not bind CR2.
Core Innovation
The invention describes antibodies and compositions useful for detection, diagnosis, and treatment of Epstein Barr Virus (EBV) infection in mammals, alongside methods of using these compositions. Specific proteins referred to as anti-gp350 antibody probes and anti-gp350 B-cell probes are also disclosed, which maintain the epitope structure of the CR2-binding region of gp350 but do not bind CR2.
The problem addressed arises from the widespread infection of EBV worldwide, often leading to serious conditions like infectious mononucleosis (IM), chronic active EBV infection (CAEBV), Hodgkin disease, lymphomas, multiple sclerosis, nasopharyngeal carcinoma, gastric carcinoma, and post-transplant lymphoproliferative disorder (PTLD). Notably, immunocompromised transplant recipients who are EBV-seronegative are at high risk of developing life-threatening PTLD because of impaired T-cell immunity. Current approaches lack effective antibody-based agents for diagnosing and treating EBV infection, highlighting the need for specific antibodies and compositions that specifically recognize EBV to reduce high risks in vulnerable patient groups.
The invention focuses on monoclonal antibodies (mAbs) to EBV gp350 protein, which initiates infection by binding to B cell receptor CR2. These mAbs were isolated from macaques immunized with a gp350 nanoparticle vaccine and characterized to be EBV-specific with up to 100-fold greater neutralizing activity compared to existing antibodies like 72A1. They recognize both cell-associated and secreted native gp350, making them valuable for immunoassay development and immunodiagnostic reagents, as well as for treatment or prevention of EBV-associated diseases.
Claims Coverage
The claims include one independent claim focusing on a modified EBV gp350 protein probe, covering multiple inventive features related to specific amino acid substitutions that reduce CR2 binding while maintaining binding to anti-gp350 antibodies and B-cells.
Modified EBV gp350 protein with reduced CR2 binding
A protein comprising a polypeptide sequence at least 85% identical to the CR2-binding region of EBV gp350, specifically residues 2-425 of SEQ ID NO:124, wherein an amino acid residue corresponding to D296 is substituted with arginine, resulting in reduced binding to CR2 but retaining the ability to bind anti-gp350 antibodies or B-cells expressing anti-gp350 BCR.
Additional amino acid substitutions on the gp350 CR2-binding region
Substitutions at positions corresponding to Q122, P158, I160, K161, W162, D163, N164 in the gp350 CR2-binding region, with specific residues given (such as substitutions to arginine, alanine, asparagine, or other amino acids) to create variants that lack CR2 binding but maintain binding to anti-gp350 antibodies or B-cells.
Insertion of amino acid residues in the CR2-binding region
Insertion of amino acid residues such as serine, threonine, cysteine, proline, asparagine, or glutamine between residues corresponding to N164 and C165 of SEQ ID NO:124 to reduce or eliminate CR2 binding while preserving antibody or B-cell receptor recognition.
Proteins comprising specific amino acid sequences
Proteins comprising amino acid sequences selected from specified SEQ ID NOs (e.g., SEQ ID NO:131, 135, 137, 139, 141, 149, 153, 155, 157, 159) representing CR2-binding region variants with reduced CR2 affinity but preserved antibody/B-cell binding.
Nucleic acids encoding the modified proteins
Nucleic acid molecules encoding the modified EBV gp350 protein variants with reduced CR2 binding as described in the claims, enabling recombinant production of the probes.
Methods of identifying anti-gp350 antibodies or anti-gp350 B-cells
Using the disclosed modified EBV gp350 protein probes to contact solutions comprising antibodies or B-cells and isolating those that specifically bind to the probes, thereby identifying anti-gp350 antibodies or B-cells.
The claims cover proteins comprising specific substitutions in the EBV gp350 CR2-binding region to eliminate or reduce CR2 binding while maintaining binding to anti-gp350 antibodies and B-cells, nucleic acids encoding these proteins, and methods for identifying corresponding antibodies or B-cells using these proteins as probes.
Stated Advantages
Monoclonal antibodies described have up to 100-fold greater EBV neutralizing activity than the most potent previously reported antibody (72A1).
These antibodies can recognize both cell-associated and secreted forms of native gp350, making them useful for both immunoassay development and clinical diagnostic confirmation of EBV infection.
The antibodies have potential for therapeutic use in treatment or prevention of EBV-associated diseases and disorders.
Documented Applications
Use as therapeutic agents for prophylaxis to prevent EBV infection in EBV-naïve transplantation recipients to reduce risk of lymphoproliferative disease.
Treatment of infectious mononucleosis or prevention of viral reactivation in persistently EBV infected individuals.
Use in diagnostic assays for detection and staging of EBV infection.
Use in purification or immunoprecipitation of EBV gp350 protein for research or clinical applications.
Use as immunodiagnostic reagents and in point-of-care lateral flow assay devices for detection and diagnosis of EBV infection.
Identification and isolation of anti-gp350 antibodies or anti-gp350 B-cells using modified gp350 protein probes.
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