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Evaluation of monoclonal antibodies for identification of subpopulations of myeloid cells in bone marrow obtained from dogs

Douglas J. WeissDepartment of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St. Paul, MN 55108.

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 DVM, PhD

Abstract

Objective—To evaluate monoclonal antibodies that may be useful for immunophenotyping myeloid cells in bone marrow of dogs.

Sample Population—Bone marrow specimens obtained from 5 dogs.

Design—Specimens were labeled with monoclonal antibodies that detected CD18, major histocompatability antigen class-II (MHC class-II), CD14, and Thy-1. Cells labeled with each of the antibodies were isolated by use of a fluorescence-activated cell sorter. Differential cell counts of sorted cells were used to determine cells that were labeled by each of the various antibodies.

Results—Myeloid cells labeled with anti-CD18 antibody included granulocytes, lymphocytes, and monocytes- macrophages. Immature and mature granulocytes were labeled. Lymphocytes, monocytesmacrophages, and eosinophils were labeled with anti-Thy-1 antibody. Cells labeled with anti-MHC-class II antibody included approximately 9% of bone marrow cells, which consisted almost exclusively of lymphocytes and monocytes-macrophages. Approximately 4% of bone marrow cells were labeled with anti-CD14 antibody, with > 90% of sorted cells being monocytes-macrophages.

Conclusions and Clinical Relevance—Four monoclonal antibodies for use in detecting subpopulations of canine bone marrow cells were evaluated. These antibodies should be useful in differentiating the origin of leukemic cells in dogs. (Am J Vet Res 2001;62:1229–1233)

Abstract

Objective—To evaluate monoclonal antibodies that may be useful for immunophenotyping myeloid cells in bone marrow of dogs.

Sample Population—Bone marrow specimens obtained from 5 dogs.

Design—Specimens were labeled with monoclonal antibodies that detected CD18, major histocompatability antigen class-II (MHC class-II), CD14, and Thy-1. Cells labeled with each of the antibodies were isolated by use of a fluorescence-activated cell sorter. Differential cell counts of sorted cells were used to determine cells that were labeled by each of the various antibodies.

Results—Myeloid cells labeled with anti-CD18 antibody included granulocytes, lymphocytes, and monocytes- macrophages. Immature and mature granulocytes were labeled. Lymphocytes, monocytesmacrophages, and eosinophils were labeled with anti-Thy-1 antibody. Cells labeled with anti-MHC-class II antibody included approximately 9% of bone marrow cells, which consisted almost exclusively of lymphocytes and monocytes-macrophages. Approximately 4% of bone marrow cells were labeled with anti-CD14 antibody, with > 90% of sorted cells being monocytes-macrophages.

Conclusions and Clinical Relevance—Four monoclonal antibodies for use in detecting subpopulations of canine bone marrow cells were evaluated. These antibodies should be useful in differentiating the origin of leukemic cells in dogs. (Am J Vet Res 2001;62:1229–1233)