Use of monoclonal antibodies to refine flow cytometric differential cell counting of canine bone marrow cells.
OBJECTIVE
To evaluate use of monoclonal antibodies to increase accuracy of flow cytometric differential cell counting of canine bone marrow cells.


METHODS
Bone marrow specimens from 15 dogs.


METHODS
Specimens were labeled with monoclonal antibodies that detected CD18, major histocompatability antigen class-II (MHC class-II), CD14, and Thy-1. Location of fluorescent and nonfluorescent cells within gates of a template developed for canine bone marrow differential cell counting was determined, the template was revised, and 10 specimens were analyzed by use of the old and revised templates and by labeling cells with anti-MHC class-II and anti-CD14.


RESULTS
Data confirmed the presumptive location of marrow subpopulations in scatter plots, permitted detection of lymphocytes and monocytemacrophages, and was used to revise the analysis template used for differential cell counting. When differential cells counts determined by the original and revised templates were compared with results of manual differential cell counts, the revised template had higher correlation coefficients and more similar mean values. Labeling cells with anti-MHC class-II and anti-CD14 permitted identification of lymphoid and monocyte-macrophages cells in bone marrow specimens.


CONCLUSIONS
Use of the revised flow cytometric analysis template combined with anti-CD14 and anti-MHC class-II antibody labeling provides reliable differential cell counts for clinical bone marrow specimens in dogs. These techniques have potential applications to clinical bone marrow examination and preclinical toxicity studies.
Weiss
D J
DJ
Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St. Paul 55108, USA.
D000818:Animals; D000911:Antibodies, Monoclonal; D001854:Bone Marrow Cells / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D018821:CD18 Antigens / Q000032:analysis; D004283:Dog Diseases / Q000276:immunology / Q000473:pathology; D004285:Dogs / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods / Q000662:veterinary*; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008285:Major Histocompatibility Complex / Q000276:immunology; D018800:Thy-1 Antigens / Q000032:analysis
D000911:Antibodies, Monoclonal; D018821:CD18 Antigens; D018950:Lipopolysaccharide Receptors; D018800:Thy-1 Antigens