Establishment and Characterization of Monoclonal Antibody Against Canine CD8 Alpha

Monoclon Antib Immunodiagn Immunother. 2020 Aug;39(4):129-134. doi: 10.1089/mab.2020.0002. Epub 2020 Jun 16.

Abstract

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.

Keywords: CD8; canine; immunohistochemistry; monoclonal antibody.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • CD8 Antigens / genetics
  • CD8 Antigens / immunology*
  • Cats
  • Cell Line
  • Dogs
  • Flow Cytometry
  • Immunohistochemistry / methods*
  • Leukocytes, Mononuclear / immunology
  • Mice
  • NIH 3T3 Cells
  • Paraffin Embedding
  • Rats

Substances

  • Antibodies, Monoclonal
  • CD8 Antigens
  • CD8 antigen, alpha chain