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Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function.

  • Author(s): Choi, Jin Wook
  • Withers, Sita S
  • Chang, Hong
  • Spanier, Justin A
  • De La Trinidad, Victoria L
  • Panesar, Harmanpreet
  • Fife, Brian T
  • Sciammas, Roger
  • Sparger, Ellen E
  • Moore, Peter F
  • Kent, Michael S
  • Rebhun, Robert B
  • McSorley, Stephen J
  • et al.
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.

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