Short E C, Fulton R W
Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Stillwater 74078.
J Clin Microbiol. 1987 Sep;25(9):1735-40. doi: 10.1128/jcm.25.9.1735-1740.1987.
2',5'-Oligoadenylate [2',5'-oligo(A)] was separated from 14C-labeled nucleosides produced in the 2',5'-oligo(A) synthetase assay by using 100-microliters columns of Dowex 1. No detectable nucleoside remained on the column after elution with 20 column volumes of water, whereas less than 1% of oligonucleotides were eluted from the column. At least 99% of oligonucleotides were eluted from the column with 1 M NaCl, pH 2. The major product had properties consistent with 2',5'-oligo(A). Exposure to alpha-1 bovine interferon (IFN) caused an increase in cellular 2',5'-oligo(A) synthetase activity which was proportional to the concentration of IFN in the medium up to 10(4) U of IFN per ml and then leveled off at about 15 X control activity. Under the assay conditions used, 2',5'-oligo(A) synthetase activity was directly proportional to the amount of cell extract over a 10-fold range. Cattle inoculated with IBR/BVD/PI-3 modified live virus vaccine showed an increase in 2',5'-oligo(A) synthetase activity in peripheral blood mononuclear leukocytes which persisted for at least 3 days postvaccination. Intramuscular injection of cattle with IFN caused a similar increase in 2',5'-oligo(A) synthetase activity. Changes in 2',5'-oligo(A) synthetase activity should be of value in (i) assessing the response of cattle to experimental viral infections or inoculations with viral vaccines or IFN or (ii) indicating a possible viral etiology in disease.
通过使用100微升的Dowex 1柱,从2',5'-寡腺苷酸合成酶测定中产生的14C标记核苷中分离出2',5'-寡腺苷酸[2',5'-oligo(A)]。用20柱体积的水洗脱后,柱上没有可检测到的核苷残留,而从柱上洗脱的寡核苷酸不到1%。至少99%的寡核苷酸用1 M NaCl,pH 2从柱上洗脱。主要产物的性质与2',5'-oligo(A)一致。暴露于α-1牛干扰素(IFN)会导致细胞2',5'-寡腺苷酸合成酶活性增加,该活性与培养基中IFN的浓度成正比,直至每毫升10(4) U的IFN,然后在约15倍对照活性时趋于平稳。在所使用的测定条件下,2',5'-寡腺苷酸合成酶活性在10倍范围内与细胞提取物的量成正比。接种IBR/BVD/PI-3改良活病毒疫苗的牛外周血单核白细胞中的2',5'-寡腺苷酸合成酶活性增加,该活性在接种疫苗后至少持续3天。给牛肌肉注射IFN会导致2',5'-寡腺苷酸合成酶活性出现类似增加。2',5'-寡腺苷酸合成酶活性的变化在以下方面应该具有价值:(i)评估牛对实验性病毒感染或接种病毒疫苗、IFN的反应,或(ii)表明疾病中可能的病毒病因。