A monoclonal antibody against a canine CD45 homologue: Analysis of tissue distribution, biochemical properties and in vitro immunological activity

Abstract

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.

Chemokine receptor 7 (CCR7)-expression and IFNγ production define vaccine-specific canine T-cell subsets

“…Clusters of antibodies collected from several sources identified canine equivalents of CD4, CD8, and Thy1.1 antigens from peripheral blood. Additional antibodies reactive to canine leukocyte antigens including CD45R (Aguiar et al, 2005) CD45RA (Caniatti et al, 1996), CD11/CD18 (Danilenko et al, 1992a; Moore et al, 1990), and CD62L (Crockett-Torabi and Fantone, 1997) and to platelet and erythrocyte antigens (Schuberth et al, 2007) have been described separately from the CLAW workshop. Testing of monoclonal antibodies specific for cytokines in other species have also identified IL-4-, IL-8-, and IFN-γ-producing canine PBMCs and expanded the repertoire of canine specific reagents (Pedersen et al, 2002).…”