OMIP-121: Immune Phenotyping of Canine Peripheral Leukocytes by Mass Cytometry

Abstract

Dogs are a critical non-rodent species used in preclinical safety studies, particularly, in the pharmaceutical field, due to their physiological, metabolic, and immunological similarities to humans. As such, immunophenotyping of canine peripheral blood mononuclear cells (PBMCs) plays a crucial role in translational research, immune monitoring, and safety evaluations in drug development. However, the limited availability of canine-specific antibodies restricts detailed and accurate immune profiling, which is essential for advancing safety evaluations in drug development. To address this challenge, we developed a 15-marker panel for comprehensive mass cytometry-based immunophenotyping of cryopreserved canine PBMCs. This panel encompasses major leukocyte subsets, including B cells, CD4+ T helper cells, regulatory T cells, CD8+ cytotoxic T cells, memory T cell subsets, natural killer T cells, natural killer cells, dendritic cells, CD4+ monocytes, classical monocytes, and neutrophils. We utilized both extracellular and intracellular markers to facilitate in-depth immune profiling, despite the limited availability of canine-specific antibodies. The panel was thoroughly optimized in terms of marker selection, antibody clone validation, and metal isotope pairing. Additionally, by the use of mass cytometry, several channels remain unoccupied, providing flexibility for future panel expansion.