The antibody against human CD25, ACT-1, recognizes canine T-lymphocytes in the G2/M and G0/G1 phases of the cell cycle during proliferation.
Because of the lack of an appropriate antibody against the canine CD25 molecule, we investigated whether anti-human CD25 antibody, ACT-1, could be useful in detecting canine T-lymphocyte proliferation. Peripheral mononuclear cells from a dog were cultured for 4 days with or without concanavalin A stimulation. In the last 24 hr, bromodeoxyuridine (BrdU) and human recombinant IL-2 were added. While the cell cycle was detected using anti-BrdU antibody and 7-amino-actinomycine (7-AAD), the cultured cells were stained with anti-canine CD4 antibody and ACT-1. The results showed that T-lymphocytes reactive to ACT-1 were present in the G2/M and G0/G1 phases in 94.4% and 70.0% of CD4-positive T-lymphocytes, respectively, suggesting that flow cytometory with ACT-1 might be useful in detecting canine T-lymphocytes during and after activation.
Masuda
Kenichi
K
Animal Allergy Clinical Laboratories Inc, Kanagawa, Japan. masuda@aacl.co.jp
Yasuda
Nobutaka
N
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D002453:Cell Cycle; D002455:Cell Division; D004285:Dogs; D016193:G1 Phase; D016195:G2 Phase; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology*
D000906:Antibodies; D015703:Antigens, CD; D053645:Interleukin-2 Receptor alpha Subunit