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CD20 Expression in Normal Canine B Cells and in Canine non-Hodgkin Lymphoma

Integrated Department of Immunology (CMJ, SPF, DC, DB, JFM), and AMC Cancer Center (CMJ, SPF, JFM), University of Colorado Health Sciences Center, Denver, CO; IHC Services (JWW), Smithville, TX; Department of Pathology and Infectious Diseases (VEOV), College of Veterinary Medicine, University of Illinois, Urbana, IL; Idexx Veterinary Services (DMG), Broomfield, CO; University of Colorado Cancer Center (DB, JFM), Denver, CO

We examined the expression of CD20 in normal canine peripheral blood mononuclear cells, normal canine spleen, and canine non-Hodgkin lymphoma (NHL) to determine the feasibility of using this antigen as a diagnostic aid and as a possible target for therapy. An antibody generated against a C-terminal (intracytoplasmic) epitope of human CD20 recognized proteins of 32–36 kd in normal and malignant canine lymphocytes. This antibody showed restricted membrane binding in a subset of lymphocytes in peripheral blood, in the B-cell regions from a normal canine spleen and lymph node, and in malignant cells from 19 dogs with B-cell NHL, but not from 15 dogs with T-cell NHL. The patterns of CD20 reactivity in these samples overlapped those seen using an antibody that recognizes canine CD79a. This anti-CD20 antibody is therefore suitable as an aid to phenotype canine NHL. In contrast, normal canine B cells were not recognized by any of 28 antibodies directed against the extracellular domains of human CD20 (including the chimeric mouse-human antibody Rituximab) or by any of 12 antibodies directed against the extracellular domains of mouse CD20. Thus, the use of CD20 as a therapeutic target will require the generation of specific antibodies against the extracellular domains of canine CD20.

Key words: B cells; canine; CD20; flow cytometry; immunoblotting; immunohistology; non-Hodgkin lymphoma.

Request reprints from Dr. Jaime F. Modiano, Integrated Department of Immunology and AMC Cancer Center, University of Colorado Health Sciences Center, 1600 Pierce Street, 2-Diamond Research Building, Denver, CO 80214 (USA). E-mail: modianoj{at}amc.org

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