Journal Article

Isolation of a monoclonal antibody from a phage display library binding the rhesus macaque MHC class I allomorph Mamu-A1001

Michaels, Anthony J., Reynolds, Matthew R., Reimann, Keith A., DeMars, Robert I., Harsla, Trevor R., Belli, Aaron J., Weinfurter, Jason T., Wiseman, Roger W., Holman, Nathan

2017

Overview

Monoclonal antibodies that bind to human leukocyte antigen (HLA) are useful tools for HLA-typing, tracking donor-recipient chimerisms after bone marrow transplants, and characterizing specific major histocompatibility complexes (MHC) on cell surfaces. Unfortunately, equivalent reagents are not available for rhesus macaques, which are commonly used animal as models in organ transplant and infectious disease research. To address this deficiency, we isolated an antibody that recognizes the common Indian rhesus macaque MHC class I molecule, Mamu-A1*001. We induced Mamu-A1*001-binding antibodies by alloimmunizing a female Mamu-A1*001-negative rhesus macaque with peripheral blood mononuclear cells (PBMC) from a male Mamu-A1*001-positive donor. A Fab phage display library was constructed with PBMC from the alloimmunized macaque and panned to isolate an antibody that binds to Mamu-A1*001 but not to other common rhesus macaque MHC class I molecules. The isolated antibody distinguishes PBMC from Mamu-A1*001-positive and -negative macaques. Additionally, the Mamu-A1*001-specific antibody binds the cynomolgus macaque MHC class I ortholog Mafa-A1*001:01 but not variants Mafa-A1*001:02/03, indicating a high degree of binding specificity. The Mamu-A1*001-specific antibody will be useful for identifying Mamu-A1*001-positive rhesus macaques, for detecting Mamu-A1*001-positive cells in populations of Mamu-A1*001-negative cells, and for examining disease processes that alter expression of Mamu-A1*001 on cell surfaces. Moreover, the alloimmunization process we describe will be useful for isolating additional MHC allomorph-specific monoclonal antibodies or antibodies against other polymorphic host proteins which are difficult to isolate with traditional technologies.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Animal models / Animals / Antibodies, Monoclonal - immunology / Antibodies, Monoclonal - isolation & purification / Antigens / B cells / Binding / Biology and life sciences / Bone marrow / Bone marrow transplantation / Cytotoxicity / Fab / Female / Histocompatibility antigen HLA / Histocompatibility Antigens Class I - immunology / HIV / HLA antigens / Human immunodeficiency virus / Humans / Immunization / Immunoglobulins / Infectious diseases / Isoimmunization / Leukocytes (mononuclear) / Lymphocytes / Macaca mulatta / Major histocompatibility complex / Male / Medical research / Medicine and Health Sciences / Monkeys & apes / Monoclonal antibodies / Pathology / Peptide Library / Peptides / Peripheral blood mononuclear cells / Phage display / Proteins / Reagents / Research and Analysis Methods / Stem cells / Tissue typing / Transplants / Transplants & implants / Viral infections