Related Experiment Videos
Characterization of dog interleukin-2 activity
S Mizuno1, T Fujinaga, M Hagio
1Department of Veterinary Surgery, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Proliferative activity of murine interleukin-2 (IL-2) dependent T cells (CTLL-2) was detected in the culture supernatant of canine peripheral blood lymphocytes (PBL) stimulated with phytohemagglutinin-P (PHA-P), and was defined as dog IL-2. The highest production of IL-2 was obtained under the conditions in which a PBL population of 2 x 10(6) cells/ml was stimulated with PHA-P at a concentration of 10 micrograms/ml at 38 degrees C for 48 hr. Dog IL-2 activity was significantly inhibited by heating at 65 degrees C, acidification under pH 4, alkalification over pH 10, and trypsin exposure. A peak of dog IL-2 activity was detected in the fraction with a molecular weight of approximately 31,000 by gel filtration. Long-term culture of canine lymphocytes was successful over 10 passages by using dog IL-2 with PHA-P-stimulation every 3 passages. The cultured cells mostly consisted of small- and medium-sized lymphocytes. These cells reacted to anti-dog thymocyte rabbit serum and anti-dog Thy-1 mouse monoclonal antibody. These cells were therefore considered to originate in T-lineage lymphocytes. Cytostasis of PBL from intact dogs reacting to canine transmissible venereal sarcoma cells was increased significantly when PBL was cultured for more than 30 days with homologous IL-2.
Proliferative activity of murine interleukin-2 (IL-2) dependent T cells (CTLL-2) was detected in the culture supernatant of canine peripheral blood lymphocytes (PBL) stimulated with phytohemagglutinin-P (PHA-P), and was defined as dog IL-2. The highest production of IL-2 was obtained under the conditions in which a PBL population of 2 x 10(6) cells/ml was stimulated with PHA-P at a concentration of 10 micrograms/ml at 38 degrees C for 48 hr. Dog IL-2 activity was significantly inhibited by heating at 65 degrees C, acidification under pH 4, alkalification over pH 10, and trypsin exposure. A peak of dog IL-2 activity was detected in the fraction with a molecular weight of approximately 31,000 by gel filtration. Long-term culture of canine lymphocytes was successful over 10 passages by using dog IL-2 with PHA-P-stimulation every 3 passages. The cultured cells mostly consisted of small- and medium-sized lymphocytes. These cells reacted to anti-dog thymocyte rabbit serum and anti-dog Thy-1 mouse monoclonal antibody. These cells were therefore considered to originate in T-lineage lymphocytes. Cytostasis of PBL from intact dogs reacting to canine transmissible venereal sarcoma cells was increased significantly when PBL was cultured for more than 30 days with homologous IL-2.