Identification and characterization of a mouse monoclonal antibody (M10) directed against canine (dog) CD8+ lymphocytes.
Monoclonal antibodies (mAb) were produced by immunizing BALB/c mice with non-adherent dog lymphocytes. M10 was specific for a subset of dog lymphocytes. M10 belonged to the IgG1 subclass and reacted with 26% of dog peripheral blood lymphocytes, 24% of spleen lymphocytes, 81% of thymus cells, 1.2% of bone marrow cells (5.8% of bone marrow lymphocytes) and 23% of PHA-stimulated lymphocytes. Immunohistology of snap-frozen thymus and spleen showed that the spleen B-cell area stained negative, whereas the spleen T-cell area and the thymus medulla exhibited positive reaction in 20-30%. The thymus cortex was strongly positive. M10 diminished cell lysis by 58% in cell mediated lysis assays (CML). Immunoblot assays revealed that M10 recognized an antigen with a molecular weight of 76 kD under non-reducing and 33 kD under reducing conditions. Finally, M10 bound to a canine CD8 alpha transfected rat T-cell line (NB2). These findings characterize M10 as an antibody directed against the dog CD8 antigen.
Voss
C
C
GSF, Institute of Immunology, Munich, Germany.
Kremmer
E
E
Hoffmann-Fezer
G
G
Schumm
M
M
Günther
W
W
Kolb
H J
HJ
Thierfelder
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D001402:B-Lymphocytes / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology*; D002460:Cell Line; D004285:Dogs / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D005434:Flow Cytometry / Q000662:veterinary; D007074:Immunoglobulin G / Q000302:isolation & purification; D007132:Immunoglobulin Isotypes / Q000302:isolation & purification; D008213:Lymphocyte Activation / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed / Q000662:veterinary; D008221:Lymphoid Tissue / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D014162:Transfection
D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes