Expression of adhesion and Fcgamma-receptors on canine blood eosinophils and neutrophils studied by anti-human monoclonal antibodies.
Commercially available anti-human monoclonal antibodies were tested as markers of adhesion receptors and Fcgamma-receptors on canine neutrophils and eosinophils. Purified populations of eosinophils and neutrophils were incubated with selected monoclonal antibodies and binding was measured by flow cytometry. Most of the anti-human monoclonal antibodies used in this study crossreacted with canine granulocytes and many showed expression similar to that of human granulocytes. The results suggest that the adhesion and Fcgamma-receptors are well conserved among species. Canine eosinophils and neutrophils were simultaneously purified by a two-layer Percoll density gradient centrifugation. The purity of the neutrophil fraction was > or = 97% after lysis of the erythrocytes. In the eosinophil fraction, 27-92% were eosinophils (mean 60%). After purification, the neutrophils and eosinophils were incubated with selected monoclonal antibodies and analysed by flow cytometry. Human anti-CD11b and CD18 antibodies bound intensely to both canine eosinophils and neutrophils. Canine neutrophils did label with anti-CD29, but eosinophils did not. Anti-CD49d bound weakly to both eosinophils and neutrophils. Both anti-CD16 and anti-CD32antibodies labelled neutrophils, but not eosinophils. There was no binding of anti-CD9 to canine neutrophils and the binding of anti-CD9 to canine eosinophils varied between dogs.
Lilliehöök
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Department of Clinical Chemistry, Swedish University of Agricultural Sciences, Uppsala. inger.lilliehook@klke.slu.se
Johannisson
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Håkansson
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D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000378:metabolism; D018821:CD18 Antigens / Q000378:metabolism; D002469:Cell Separation; D003429:Cross Reactions; D004285:Dogs / Q000097:blood* / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D039441:Integrin alpha4; D019012:Integrin beta1 / Q000378:metabolism; D016023:Integrins / Q000097:blood* / Q000276:immunology; D016177:Macrophage-1 Antigen / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins; D017452:Receptors, IgG / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D013045:Species Specificity; D060245:Tetraspanin 29
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018821:CD18 Antigens; C554869:CD9 protein, human; D019012:Integrin beta1; D016023:Integrins; D016177:Macrophage-1 Antigen; D008562:Membrane Glycoproteins; D017452:Receptors, IgG; D060245:Tetraspanin 29; D039441:Integrin alpha4