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Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function
Jin Wook Choi1, Sita S Withers2, Hong Chang2
1Center for Immunology and Infectious Diseases, Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, California, United States of America.
Abstract:
Interruption of the programmed death 1 (PD-1) / programmed death ligand 1 (PD-L1) pathway is an established and effective therapeutic strategy in human oncology and holds promise for veterinary oncology. We report the generation and characterization of monoclonal antibodies specific for canine PD-1 and PD-L1. Antibodies were initially assessed for their capacity to block the binding of recombinant canine PD-1 to recombinant canine PD-L1 and then ranked based on efficiency of binding as judged by flow cytometry. Selected antibodies were capable of detecting PD-1 and PD-L1 on canine tissues by flow cytometry and Western blot. Anti-PD-L1 worked for immunocytochemistry and anti-PD-1 worked for immunohistochemistry on formalin-fixed paraffin embedded canine tissues, suggesting the usage of this antibody with archived tissues. Additionally, anti-PD-L1 (JC071) revealed significantly increased PD-L1 expression on canine monocytes after stimulation with peptidoglycan or lipopolysaccharide. Together, these antibodies display specificity for the natural canine ligand using a variety of potential diagnostic applications. Importantly, multiple PD-L1-specific antibodies amplified IFN-γ production in a canine peripheral blood mononuclear cells (PBMC) concanavlin A (Con A) stimulation assay, demonstrating functional activity.
Insights
Researchers developed novel antibodies targeting canine programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1). These tools show promise for diagnosing and treating cancer in dogs by blocking the PD-1/PD-L1 pathway.
Area of Science:
- Veterinary Oncology
- Immunology
- Monoclonal Antibody Development
Background:
- The programmed death 1 (PD-1) / programmed death ligand 1 (PD-L1) pathway is a validated therapeutic target in human cancer treatment.
- This pathway's potential in veterinary oncology necessitates the development of specific reagents for canine applications.
Purpose of the Study:
- To generate and characterize monoclonal antibodies targeting canine PD-1 and PD-L1.
- To evaluate the diagnostic and functional potential of these antibodies in canine samples.
Main Methods:
- Generation of monoclonal antibodies against canine PD-1 and PD-L1.
- Assessment of antibody binding affinity and specificity using flow cytometry and Western blot.
- Validation of antibody utility in immunocytochemistry, immunohistochemistry, and functional assays (IFN-γ production).
Main Results:
- Successfully generated and characterized canine PD-1 and PD-L1 specific monoclonal antibodies.
- Antibodies demonstrated effective detection of target proteins in canine tissues and cells.
- Anti-PD-L1 antibody (JC071) showed increased expression on canine monocytes post-stimulation.
- Functional assays confirmed that PD-L1 antibodies can enhance IFN-γ production in canine PBMCs.
Conclusions:
- The developed antibodies are specific for canine PD-1 and PD-L1, suitable for diagnostic applications.
- These reagents hold significant promise for advancing immunotherapy research and treatment strategies in veterinary oncology.
- The antibodies' ability to detect target proteins in formalin-fixed tissues expands their utility for retrospective studies.
