Swine interleukin 2 activity produced by mesenteric lymph node cells.
Swine interleukin 2 (IL-2) activity was assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay using murine IL-2 dependent cell line (CTLL-2). The culture supernatant of mesenteric lymph node (MLN) cells stimulated with phytohemagglutinin-P (PHA) induced a generation of MTT formazan in a dose-dependent manner, suggesting dose-dependent proliferation of CTLL-2. The maximal swine IL-2 activity was observed in the culture of MLN cells at 1 to 2 x 10(7) cells/ml when stimulated with 20 to 40 micrograms/ml PHA for 48 hr. Based on these findings, a large culture of MLN cells to prepare swine IL-2 were performed under the following condition; cell concentration of 1 x 10(7) cells/ml, PHA concentration of 20 micrograms/ml, a culture scale of 200 to 400 ml, and a stirring speed of 30 rpm. Swine IL-2 activity was detected from 4 hr after PHA stimulation, and rapidly increased until 16 hr. Almost maximal IL-2 activity in stirring culture was observed at the incubation of 20 hr. Swine IL-2 was partially purified by Sephacryl S-200 gel filtration and the estimated molecular weight was about 32 kD based on the peak of IL-2 activity. The pI value of swine IL-2 was estimated to be approximately pH 5.3. Swine IL-2 was sensitive to acid (pH 3.2) or alkaline (pH 10.5), 4 or 8 M urea, trypsin, and the heating at 70 degrees C. These physico-chemical properties of swine IL-2 was similar to those of human, murine or feline IL-2.
Iwata
H
H
Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Japan.
Ueda
T
T
Takayanagi
K
K
Wada
M
M
Inoue
T
T
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D007376:Interleukin-2 / Q000032:analysis / Q000096:biosynthesis*; D007700:Kinetics; D008198:Lymph Nodes / Q000276:immunology / Q000378:metabolism*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D013154:Spleen / Q000276:immunology; D013552:Swine
D007376:Interleukin-2