Understanding of the microenvironment of cancer plays a crucial role in cancer research. A tool is needed to evaluate the immune cells surrounding the cancer cells. This study establishes and evaluates a novel monoclonal antibody against canine CD8α (cCD8α). The antibody was produced by immunization of rats with cCD8α-expressing cells. After establishment and selection of hybridoma cells, the clone F3-B2 was established. The reactivity of F3-B2 was confirmed using cCD8α-overexpressing murine cells. Flow cytometric analysis also demonstrated that F3-B2 reacts with cCD8α naturally expressed in canine peripheral blood mononuclear cells and a canine T cell lymphoma cell line. The specimens of lymphoid tissue showed immunohistochemical staining for F3-B2. Moreover, we also found that F3-B2 exhibited reactivity against feline CD8. Thus, this antibody provides a good research tool to analyze CD8-positive cytotoxic lymphocytes in canine and feline tumors.
