Characterization of a novel bovine leukocyte protein involved in cell-cell adhesion.
Preliminary characterization of an apparently novel bovine leukocyte adhesion protein is described. Two IgG1 monoclonal antibodies, UC-C1 and UC-H5, raised against established cultures of IL-2-dependent bovine peripheral blood lymphocytes (PBL) were found to react with an antigen expressed by the majority of bovine peripheral blood leukocytes. Immunoprecipitation and polyacrylamide gel electrophoresis of the antigen produced a distinct protein band of molecular weight 160,000, and additional diffuse protein bands of approximate molecular weight 180,000, 175,000, and 150,000. Two-color flow cytometric analyses showed that the antigen was expressed at low density on a small proportion of circulating B lymphocytes, but was highly expressed on all circulating T lymphocytes. The majority of monocytes and all granulocytes expressed the antigen at a density lower than that of T lymphocytes. Peripheral blood lymphocytes stimulated with concanavalin A had an approximately 3-fold increased expression of the antigen, which was apparent within 18 h and remained stable in long-term cultures. Expression of the antigen in thymus, analyzed by the immunoperoxidase technique, was predominantly restricted to thymocytes in the immediate subcapsular cortex and medulla; expression in lymph nodes and spleen was predominantly confined to lymphocytes in T-cell areas. Flow-cytometric analysis demonstrated that thymocytes and the majority of peripheral and mesenteric lymph node-derived T cells had relatively low surface density of antigen compared to circulating T cells. Binding of UC-C1 or UC-H5 to the antigen on lymphocytes induced homotypic aggregation. UC-C1 completely blocked binding of FITC-conjugated UC-H5 to blood mononuclear cells, suggesting that the antibodies recognize the same epitope or proximal epitopes.
Taylor
B C
BC
Department of Veterinary Pathology, Microbiology and Immunology, University of California, Davis.
Mattapallil
J
J
Moore
P F
PF
Scibienski
R J
RJ
Stott
J L
JL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D001798:Blood Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D002417:Cattle / Q000276:immunology / Q000378:metabolism*; D002448:Cell Adhesion; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D007074:Immunoglobulin G / Q000276:immunology; D007962:Leukocytes / Q000166:cytology / Q000276:immunology*; D016131:Lymphocyte Subsets / Q000166:cytology / Q000276:immunology; D008221:Lymphoid Tissue / Q000166:cytology; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001798:Blood Proteins; D000939:Epitopes; D007074:Immunoglobulin G