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-11236151-B2
Publication Date
2022-02-01
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 infection in mammals, and to methods of using those compositions of matter for the same. Also disclosed 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 relates to 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. The invention also includes proteins termed 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.
EBV is a human herpes virus infecting over 90% of the world population, often leading to various diseases such as infectious mononucleosis, Hodgkin disease, lymphomas, multiple sclerosis, and malignancies like nasopharyngeal and gastric carcinomas. Immunocompromised patients, especially transplant recipients who are EBV-seronegative prior to transplantation, are at high risk of post-transplant lymphoproliferative disorder (PTLD) due to primary EBV infections caused by donor organ transmission and impaired T-cell immunity. These conditions highlight the need for improved diagnostic and therapeutic agents capable of detecting EBV and inhibiting EBV infection and replication.
The invention is based on isolating a variety of monoclonal antibodies (mAbs) against EBV glycoprotein 350 (gp350) from macaque monkeys immunized with a gp350 nanoparticle vaccine. The mAbs have up to 100-fold greater EBV neutralizing activity than 72A1, the most potent previously reported murine antibody targeting the CR2-binding site on gp350. These mAbs recognize both cell-associated and secreted native gp350 forms, making them valuable for immunoassay development, clinical diagnostics, and treatment or prevention of EBV-associated diseases.
Claims Coverage
The claims disclose multiple independent inventive features focusing on isolated antibodies binding EBV gp350 protein with defined variable heavy and light domain CDR sequences, methods of using these antibodies for diagnosis, treatment, and inhibition of EBV-infected cells, as well as engineered antibody variants with specific amino acid substitutions for enhanced properties.
Isolated antibodies with specific variable heavy and light chain CDR sequences
Isolated antibodies that bind EBV gp350 protein comprising variable heavy (VH) and light (VL) domains with specific complementarity determining region (CDR) sequences as set forth in various SEQ ID NOs, including engineered antibody variants.
Isolated antibodies comprising defined VH and VL domain sequences
Isolated antibodies selected from a group of clones comprising specific VH and VL domain amino acid sequences as provided by SEQ ID NOs, including B03, E04, D09, C02, H02, and others, some being engineered.
Antibody fragments, chimeric, humanized, and bispecific forms
The antibodies encompass monoclonal, chimeric, humanized, bispecific, and fragment forms capable of binding EBV gp350 protein.
Antibodies conjugated to growth inhibitory, cytotoxic agents or labels
Anti-gp350 antibodies conjugated to agents such as growth inhibitory agents, cytotoxic agents, radioisotopes, or fluorescent labels for therapeutic or diagnostic purposes.
Nucleic acid molecules encoding the isolated antibodies
Isolated nucleic acid molecules that encode the antibodies described, enabling recombinant production.
Methods of inhibiting growth of EBV gp350-expressing cells
Methods involving contacting EBV gp350-expressing cells with the antibodies to inhibit cell growth, applicable to infected cells, epithelial cells, B-lymphocytes, and cancer cells.
Methods of treating or preventing EBV infection or associated disease
Administering a therapeutically effective amount of the isolated antibodies to individuals for treatment or prevention of EBV infection or EBV-associated diseases.
Methods of diagnosing EBV infection using antibody binding
Diagnostic methods involving contacting test samples with the antibodies and detecting complexes formed with EBV gp350 protein indicative of infection.
Detectably labeled antibodies for diagnostic use
Use of antibodies detectably labeled with radioisotopes or fluorescent labels to enhance detection in diagnostic assays.
Antibodies attached to solid supports for assays or purification
Antibodies conjugated to solid supports composed of glass, polysaccharides, polyacrylamides, polystyrene, or other materials for use in diagnostic assays or purification.
Engineered antibodies with specific amino acid substitutions for potency
Antibody variants engineered with specific amino acid substitutions in VH and VL domains, guided by Kabat numbering, to improve binding affinity and neutralization potency, with exemplar substitutions detailed.
The claims comprehensively cover isolated antibodies against EBV gp350 with defined variable regions and CDRs, their nucleic acid sequences, various engineered and conjugated antibody forms, methods of inhibiting EBV gp350-expressing cells, as well as diagnostic and therapeutic methods using these antibodies. Specific engineered substitutions further enhance antibody functionality.
Stated Advantages
The disclosed monoclonal antibodies have up to 100-fold greater EBV neutralizing activity than the most potent prior antibody (72A1).
The antibodies can recognize both cell-associated and secreted forms of native gp350, facilitating immunoassay development and clinical diagnostics.
Potential for use in therapeutic treatment or prevention of EBV-associated diseases and disorders.
Anti-gp350 antibody probes and B-cell probes can bind anti-gp350 antibodies or B-cells while lacking CR2 binding, improving specificity in isolation and detection.
Documented Applications
Detection, diagnosis, and treatment of EBV infection in mammals including humans.
Use in immunoassay development for detecting EBV in clinical samples and tissues.
Therapeutic treatment or prevention of EBV associated diseases such as infectious mononucleosis, Hodgkin's lymphoma, Burkitt's lymphoma, gastric cancer, nasopharyngeal carcinoma, autoimmune diseases including dermatomyositis, systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, multiple sclerosis, and PTLD.
Isolation and analysis of antigen-specific B-cells using anti-gp350 antibody probes or B-cell probes that retain the epitope structure but do not bind CR2 receptor.
Use in prophylaxis to prevent EBV infection in EBV-naïve transplant recipients at high risk of lymphoproliferative disease.
Treatment of infectious mononucleosis and prevention of virus reactivation in persistently infected individuals.
Use in antibody-based diagnostic kits, assay devices, and lateral flow assay devices for point-of-care EBV detection.
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